Cxcr4 inhibitors and uses thereof

ABSTRACT

The present invention provides compounds, compositions thereof, and methods of using the same.

CROSS-REFERENCE TO RELATED APPLICATIONS

This application is a continuation of U.S. patent application Ser. No.16/311,020, filed Dec. 18, 2018, which is a national stage filing under35 U.S.C. § 371 of PCT International Patent ApplicationPCT/US2017/038609, filed Jun. 21, 2017, which claims the benefit under35 U.S.C. § 119(e) of U.S. Provisional Application Nos. 62/352,820,filed Jun. 21, 2016; and 62/456,526, filed Feb. 8, 2017; each of whichis hereby incorporated by reference in its entirety.

TECHNICAL FIELD OF THE INVENTION

The present invention relates to compounds and methods useful forinhibition of C-X-C receptor type 4 (CXCR4). The invention also providespharmaceutically acceptable compositions comprising compounds of thepresent invention and methods of using said compositions in thetreatment of various disorders.

BACKGROUND OF THE INVENTION

C-X-C chemokine receptor type 4 (CXCR4), also known as fusin or clusterof differentiation 184 (CD184), is a seven transmembrane G-proteincoupled receptor (GPCR) belonging to Class I GPCR or rhodopsin-like GPCRfamily. Under normal physiological conditions, CXCR4 carries outmultiple roles and is principally expressed in the hematopoietic andimmune systems. CXCR4 was initially discovered as one of theco-receptors involved in human immunodeficiency virus (HIV) cell entry.Subsequent studies showed that it is expressed in many tissues,including brain, thymus, lymphatic tissues, spleen, stomach, and smallintestine, and also specific cell types such as hematopoietic stem cells(HSC), mature lymphocytes, and fibroblasts. CXCL12, previouslydesignated SDF-1α, is the only known ligand for CXCR4. CXCR4 mediatesmigration of stem cells during embryonic development as well as inresponse to injury and inflammation. Multiple roles have beendemonstrated for CXCR4 in human diseases such as cellular proliferativedisorders, Alzheimer's disease, HIV, rheumatoid arthritis, pulmonaryfibrosis, and others. For example, expression of CXCR4 and CXCL12 havebeen noted in several tumor types. CXCL12 is expressed bycancer-associated fibroblast (CAFs) and is often present at high levelsin the tumor microenvironment (TME). In clinical studies of a wide rangeof tumor types, including breast, ovarian, renal, lung, and melanoma,expression of CXCR4/CXCL12 has been associated with a poor prognosis andwith an increased risk of metastasis to lymph nodes, lung, liver, andbrain, which are sites of CXCL12 expression. CXCR4 is frequentlyexpressed on melanoma cells, particularly the CD133+ population that isconsidered to represent melanoma stem cells; in vitro experiments andmurine models have demonstrated that CXCL12 is chemotactic for suchcells.

Furthermore, there is now evidence implicating the CXCL12/CXCR4 axis incontributing to the loss or lack of tumor responsiveness to angiogenesisinhibitors (also referred to as “angiogenic escape”). In animal cancermodels, interference with CXCR4 function has been demonstrated to alterthe TME and sensitize the tumor to immune attack by multiple mechanismssuch as elimination of tumor re-vascularization and increasing the ratioof CD8+ T cells to Treg cells. These effects result in significantlydecreased tumor burden and increased overall survival in xenograft,syngeneic, and transgenic cancer models. See Vanharanta et al. (2013)Nat Med 19: 50-56; Gale and McColl (1999) BioEssays 21: 17-28; Highfillet al. (2014) Sci Transl Med 6: ra67; Facciabene et al. (2011) Nature475: 226-230.

These data underscore the significant, unmet need for CXCR4 inhibitorsto treat the many diseases and conditions mediated by aberrant orundesired expression of the receptor, for example in cellularproliferative disorders.

SUMMARY OF THE INVENTION

It has now been found that compounds of the present invention, andpharmaceutically acceptable compositions thereof, are effective as CXCR4inhibitors. In one aspect, the present invention provides a compound ofFormula I:

or a pharmaceutically acceptable salt thereof, wherein each variable isas defined and described herein.

Compounds of the present invention, and pharmaceutically acceptablecompositions thereof, are useful for treating a variety of diseases,disorders or conditions, associated with CXC receptor type 4 (CXCR4).Such diseases, disorders, or conditions include cellular proliferativedisorders (e.g., cancer) such as those described herein.

DETAILED DESCRIPTION OF CERTAIN EMBODIMENTS 1. General Description ofCertain Embodiments of the Invention

Compounds of the present invention, and pharmaceutical compositionsthereof, are useful as inhibitors of CXCR4. Without wishing to be boundby any particular theory, it is believed that compounds of the presentinvention, and pharmaceutical compositions thereof, may inhibit theactivity of CXCR4 and thus treat certain diseases, such as cancer.

It has now been found that compounds of this invention, andpharmaceutically acceptable compositions thereof, are effective as CXCR4inhibitors. In one aspect, the present invention provides a compound ofFormula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   Ring A is a 3-8 membered saturated or partially unsaturated    monocyclic carbocyclic ring, phenyl, an 8-10 membered bicyclic    aromatic carbocyclic ring, a 4-8 membered saturated or partially    unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms    independently selected from nitrogen, oxygen, or sulfur, a 5-6    membered monocyclic heteroaromatic ring having 1-4 heteroatoms    independently selected from nitrogen, oxygen, or sulfur, or an 8-10    membered bicyclic heteroaromatic ring having 1-5 heteroatoms    independently selected from nitrogen, oxygen, or sulfur;-   each R¹ is independently —R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃,    —SR, or -L¹-R⁶;-   each R is independently hydrogen or an optionally substituted group    selected from C₁₋₆ aliphatic, a 3-8 membered saturated or partially    unsaturated monocyclic carbocyclic ring, phenyl, an 8-10 membered    bicyclic aromatic carbocyclic ring, a 4-8 membered saturated or    partially unsaturated monocyclic heterocyclic ring having 1-2    heteroatoms independently selected from nitrogen, oxygen, or sulfur,    a 5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatoms    independently selected from nitrogen, oxygen, or sulfur, or an 8-10    membered bicyclic heteroaromatic ring having 1-5 heteroatoms    independently selected from nitrogen, oxygen, or sulfur;    -   each L¹ and L² is independently a covalent bond or a C₁₋₈        bivalent straight or branched hydrocarbon chain wherein 1, 2, or        3 methylene units of the chain are independently and optionally        replaced with —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—, —(R)NC(O)—,        —OC(O)N(R)—, —(R)NC(O)O—, —N(R)C(O)N(R)—, —S—, —SO—, —SO₂N(R)—,        —(R)NSO₂—, —C(S)—, —C(S)O—, —OC(S)—, —C(S)N(R)—, —(R)NC(S)—,        —(R)NC(S)N(R)—, or -Cy-; each -Cy- is independently a bivalent        optionally substituted 3-8 membered saturated or partially        unsaturated monocyclic carbocyclic ring, optionally substituted        phenylene, an optionally substituted 4-8 membered saturated or        partially unsaturated monocyclic heterocyclic ring having 1-3        heteroatoms independently selected from nitrogen, oxygen, or        sulfur, an optionally substituted 5-6 membered monocyclic        heteroaromatic ring having 1-4 heteroatoms independently        selected from nitrogen, oxygen, or sulfur, an optionally        substituted 8-10 membered bicyclic or bridged bicyclic saturated        or partially unsaturated heterocyclic ring having 1-5        heteroatoms independently selected from nitrogen, oxygen, or        sulfur, or an optionally substituted 8-10 membered bicyclic or        bridged bicyclic heteroaromatic ring having 1-5 heteroatoms        independently selected from nitrogen, oxygen, or sulfur;-   R² is hydrogen, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃, —SR, -L²-R⁶,    or optionally substituted C₁₋₈ aliphatic;-   R³ is hydrogen, optionally substituted C₁₋₆ aliphatic, or -L³-R⁶;-   L³ is a C₁₋₆ bivalent straight or branched hydrocarbon chain wherein    1, 2, or 3 methylene units of the chain are independently and    optionally replaced with —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—,    —C(O)N(R)—, —(R)NC(O)—, —S—, —SO—, —SO₂—, —C(S)—, or -Cy-;-   each R⁴ is independently hydrogen, deuterium, halogen, —CN, —OR⁶, or    C₁₋₄ alkyl, or two R⁴ groups on the same carbon are optionally taken    together to form ═NR⁶, ═NOR⁶, ═O, or ═S;-   each R⁵ is independently R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃,    —SR, or -L¹-R⁶, or two R⁵ groups on the same saturated carbon atom    are optionally taken together to form ═NR, ═NOR, ═O, ═S, or a    spirocyclic 3-6 membered carbocyclic ring;-   each R⁶ is independently hydrogen or C₁₋₆ alkyl optionally    substituted with 1, 2, 3, 4, 5, or 6 deuterium or halogen atoms;-   m is 0, 1, 2, 3, or 4;-   n is 0, 1, 2, 3, or 4; and-   p is 0, 1, 2, 3, or 4.

2. Compounds and Definitions

Compounds of the present invention include those described generallyherein, and are further illustrated by the classes, subclasses, andspecies disclosed herein. As used herein, the following definitionsshall apply unless otherwise indicated. For purposes of this invention,the chemical elements are identified in accordance with the PeriodicTable of the Elements, CAS version, Handbook of Chemistry and Physics,75^(th) Ed. Additionally, general principles of organic chemistry aredescribed in “Organic Chemistry”, Thomas Sorrell, University ScienceBooks, Sausalito: 1999, and “March's Advanced Organic Chemistry”, 5^(th)Ed., Ed.: Smith, M. B. and March, J., John Wiley & Sons, New York: 2001,the entire contents of which are hereby incorporated by reference.

The term “aliphatic” or “aliphatic group”, as used herein, means astraight-chain (i.e., unbranched) or branched, substituted orunsubstituted hydrocarbon chain that is completely saturated or thatcontains one or more units of unsaturation, or a monocyclic hydrocarbonor bicyclic hydrocarbon that is completely saturated or that containsone or more units of unsaturation, but which is not aromatic (alsoreferred to herein as “carbocycle,” “cycloaliphatic” or “cycloalkyl”),that has a single point of attachment to the rest of the molecule.Unless otherwise specified, aliphatic groups contain 1-6 aliphaticcarbon atoms. In some embodiments, aliphatic groups contain 1-5aliphatic carbon atoms. In other embodiments, aliphatic groups contain1-4 aliphatic carbon atoms. In still other embodiments, aliphatic groupscontain 1-3 aliphatic carbon atoms, and in yet other embodiments,aliphatic groups contain 1-2 aliphatic carbon atoms. In someembodiments, “cycloaliphatic” (or “carbocycle” or “cycloalkyl”) refersto a monocyclic C3-C6 hydrocarbon that is completely saturated or thatcontains one or more units of unsaturation, but which is not aromatic,that has a single point of attachment to the rest of the molecule.Suitable aliphatic groups include, but are not limited to, linear orbranched, substituted or unsubstituted alkyl, alkenyl, alkynyl groupsand hybrids thereof such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl or(cycloalkyl)alkenyl.

As used herein, the term “bicyclic ring” or “bicyclic ring system”refers to any bicyclic ring system, i.e. carbocyclic or heterocyclic,saturated or having one or more units of unsaturation, having one ormore atoms in common between the two rings of the ring system. Thus, theterm includes any permissible ring fusion, such as ortho-fused orspirocyclic. As used herein, the term “heterobicyclic” is a subset of“bicyclic” that requires that one or more heteroatoms are present in oneor both rings of the bicycle. Such heteroatoms may be present at ringjunctions and are optionally substituted, and may be selected fromnitrogen (including N-oxides), oxygen, sulfur (including oxidized formssuch as sulfones and sulfonates), phosphorus (including oxidized formssuch as phosphates), boron, etc. In some embodiments, a bicyclic grouphas 7-12 ring members and 0-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur. As used herein, the term “bridged bicyclic”refers to any bicyclic ring system, i.e. carbocyclic or heterocyclic,saturated or partially unsaturated, having at least one bridge. Asdefined by IUPAC, a “bridge” is an unbranched chain of atoms or an atomor a valence bond connecting two bridgeheads, where a “bridgehead” isany skeletal atom of the ring system which is bonded to three or moreskeletal atoms (excluding hydrogen). In some embodiments, a bridgedbicyclic group has 7-12 ring members and 0-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur. Such bridged bicyclic groupsare well known in the art and include those groups set forth below whereeach group is attached to the rest of the molecule at any substitutablecarbon or nitrogen atom. Unless otherwise specified, a bridged bicyclicgroup is optionally substituted with one or more substituents as setforth for aliphatic groups. Additionally or alternatively, anysubstitutable nitrogen of a bridged bicyclic group is optionallysubstituted. Exemplary bicyclic rings include:

Exemplary bridged bicyclics include:

The term “lower alkyl” refers to a C₁₋₄ straight or branched alkylgroup. Exemplary lower alkyl groups are methyl, ethyl, propyl,isopropyl, butyl, isobutyl, and tert-butyl.

The term “lower haloalkyl” refers to a C₁₋₄ straight or branched alkylgroup that is substituted with one or more halogen atoms.

The term “heteroatom” means one or more of oxygen, sulfur, nitrogen,phosphorus, or silicon (including, any oxidized form of nitrogen,sulfur, phosphorus, or silicon; the quaternized form of any basicnitrogen or; a substitutable nitrogen of a heterocyclic ring, forexample N (as in 3,4-dihydro-2H-pyrrolyl), NH (as in pyrrolidinyl) orNR⁺ (as in N-substituted pyrrolidinyl)).

The term “unsaturated”, as used herein, means that a moiety has one ormore units of unsaturation.

As used herein, the term “bivalent C18 (or C₁₋₆) saturated orunsaturated, straight or branched, hydrocarbon chain”, refers tobivalent alkylene, alkenylene, and alkynylene chains that are straightor branched as defined herein.

The term “alkylene” refers to a bivalent alkyl group. An “alkylenechain” is a polymethylene group, i.e., —(CH₂)_(n)—, wherein n is apositive integer, preferably from 1 to 6, from 1 to 4, from 1 to 3, from1 to 2, or from 2 to 3. A substituted alkylene chain is a polymethylenegroup in which one or more methylene hydrogen atoms are replaced with asubstituent. Suitable substituents include those described below for asubstituted aliphatic group.

The term “alkenylene” refers to a bivalent alkenyl group. A substitutedalkenylene chain is a polymethylene group containing at least one doublebond in which one or more hydrogen atoms are replaced with asubstituent. Suitable substituents include those described below for asubstituted aliphatic group.

As used herein, the term “cyclopropylenyl” refers to a bivalentcyclopropyl group of the following structure:

The term “halogen” means F, Cl, Br, or I.

The term “aryl” used alone or as part of a larger moiety as in“aralkyl,” “aralkoxy,” or “aryloxyalkyl,” refers to monocyclic orbicyclic ring systems having a total of five to fourteen ring members,wherein at least one ring in the system is aromatic and wherein eachring in the system contains 3 to 7 ring members. The term “aryl” may beused interchangeably with the term “aryl ring.” In certain embodimentsof the present invention, “aryl” refers to an aromatic ring system whichincludes, but not limited to, phenyl, biphenyl, naphthyl, anthracyl andthe like, which may bear one or more substituents. Also included withinthe scope of the term “aryl,” as it is used herein, is a group in whichan aromatic ring is fused to one or more non-aromatic rings, such asindanyl, phthalimidyl, naphthimidyl, phenanthridinyl, ortetrahydronaphthyl, and the like.

The terms “heteroaryl” and “heteroar-,” used alone or as part of alarger moiety, e.g., “heteroaralkyl,” or “heteroaralkoxy,” refer togroups having 5 to 10 ring atoms, preferably 5, 6, or 9 ring atoms;having 6, 10, or 14 π electrons shared in a cyclic array; and having, inaddition to carbon atoms, from one to five heteroatoms. The term“heteroatom” refers to nitrogen, oxygen, or sulfur, and includes anyoxidized form of nitrogen or sulfur, and any quaternized form of a basicnitrogen. Heteroaryl groups include, without limitation, thienyl,furanyl, pyrrolyl, imidazolyl, pyrazolyl, triazolyl, tetrazolyl,oxazolyl, isoxazolyl, oxadiazolyl, thiazolyl, isothiazolyl,thiadiazolyl, pyridyl, pyridazinyl, pyrimidinyl, pyrazinyl, indolizinyl,purinyl, naphthyridinyl, and pteridinyl. The terms “heteroaryl” and“heteroar-”, as used herein, also include groups in which aheteroaromatic ring is fused to one or more aryl, cycloaliphatic, orheterocyclyl rings, where the radical or point of attachment is on theheteroaromatic ring. Nonlimiting examples include indolyl, isoindolyl,benzothienyl, benzofuranyl, dibenzofuranyl, indazolyl, benzimidazolyl,benzthiazolyl, quinolyl, isoquinolyl, cinnolinyl, phthalazinyl,quinazolinyl, quinoxalinyl, 4H-quinolizinyl, carbazolyl, acridinyl,phenazinyl, phenothiazinyl, phenoxazinyl, tetrahydroquinolinyl,tetrahydroisoquinolinyl, and pyrido[2,3-b]-1,4-oxazin-3(4H)-one. Aheteroaryl group may be mono- or bicyclic. The term “heteroaryl” may beused interchangeably with the terms “heteroaryl ring,” “heteroarylgroup,” or “heteroaromatic,” any of which terms include rings that areoptionally substituted. The term “heteroaralkyl” refers to an alkylgroup substituted by a heteroaryl, wherein the alkyl and heteroarylportions independently are optionally substituted.

As used herein, the terms “heterocycle,” “heterocyclyl,” “heterocyclicradical,” and “heterocyclic ring” are used interchangeably and refer toa stable 5- to 7-membered monocyclic or 7-10-membered bicyclicheterocyclic moiety that is either saturated or partially unsaturated,and having, in addition to carbon atoms, one or more, preferably one tofour, heteroatoms, as defined above. When used in reference to a ringatom of a heterocycle, the term “nitrogen” includes a substitutednitrogen. As an example, in a saturated or partially unsaturated ringhaving 0-3 heteroatoms selected from oxygen, sulfur or nitrogen, thenitrogen may be N (as in 3,4-dihydro-2H-pyrrolyl), NH (as inpyrrolidinyl), or ⁺NR (as in N-substituted pyrrolidinyl).

A heterocyclic ring can be attached to its pendant group at anyheteroatom or carbon atom that results in a stable structure and any ofthe ring atoms can be optionally substituted. Examples of such saturatedor partially unsaturated heterocyclic radicals include, withoutlimitation, tetrahydrofuranyl, tetrahydrothiophenyl, pyrrolidinyl,piperidinyl, pyrrolinyl, tetrahydroquinolinyl, tetrahydroisoquinolinyl,decahydroquinolinyl, oxazolidinyl, piperazinyl, dioxanyl, dioxolanyl, diazepinyl, oxazepinyl, thiazepinyl, morpholinyl, and quinuclidinyl. Theterms “heterocycle,” “heterocyclyl,” “heterocyclyl ring,” “heterocyclicgroup,” “heterocyclic moiety,” and “heterocyclic radical,” are usedinterchangeably herein, and also include groups in which a heterocyclylring is fused to one or more aryl, heteroaryl, or cycloaliphatic rings,such as indolinyl, 3H-indolyl, chromanyl, phenanthridinyl, ortetrahydroquinolinyl. A heterocyclyl group may be mono- or bicyclic. Theterm “heterocyclylalkyl” refers to an alkyl group substituted by aheterocyclyl, wherein the alkyl and heterocyclyl portions independentlyare optionally substituted.

As used herein, the term “partially unsaturated” refers to a ring moietythat includes at least one double or triple bond. The term “partiallyunsaturated” is intended to encompass rings having multiple sites ofunsaturation, but is not intended to include aryl or heteroarylmoieties, as herein defined.

As described herein, compounds of the invention may contain “optionallysubstituted” moieties. In general, the term “substituted,” whetherpreceded by the term “optionally” or not, means that one or morehydrogens of the designated moiety are replaced with a suitablesubstituent. Unless otherwise indicated, an “optionally substituted”group may have a suitable substituent at each substitutable position ofthe group, and when more than one position in any given structure may besubstituted with more than one substituent selected from a specifiedgroup, the substituent may be either the same or different at everyposition. Combinations of substituents envisioned by this invention arepreferably those that result in the formation of stable or chemicallyfeasible compounds. The term “stable,” as used herein, refers tocompounds that are not substantially altered when subjected toconditions to allow for their production, detection, and, in certainembodiments, their recovery, purification, and use for one or more ofthe purposes disclosed herein.

Each optional substituent on a substitutable carbon is a monovalentsubstituent independently selected from halogen; —(CH₂)₀₋₄R^(∘);—(CH₂)₀₋₄OR^(∘); —O(CH₂)₀₋₄R^(∘), —O—(CH₂)₀₋₄C(O)OR^(∘);—(CH₂)₀₋₄CH(OR^(∘))₂; —(CH₂)₀₋₄SR^(∘); —(CH₂)₀₋₄Ph, which may besubstituted with R^(∘); —(CH₂)₀₋₄O(CH₂)₀₋₁Ph which may be substitutedwith R^(∘); —CH═CHPh, which may be substituted with R^(∘);—(CH₂)₀₋₄O(CH₂)₀₋₁-pyridyl which may be substituted with R^(∘); —NO₂;—CN; —N₃; —(CH₂)₀₋₄N(R^(∘))₂; —(CH₂)₀₋₄N(R^(∘))C(O)R^(∘);—N(R^(∘))C(S)R^(∘); —(CH₂)₀₋₄N(R^(∘))C(O)NR^(∘))₂;—N(R^(∘))C(S)NR^(∘))₂; —(CH₂)₀₋₄N(R^(∘))C(O)OR^(∘);—N(R^(∘))N(R^(∘))C(O)R^(∘); —N(R^(∘))N(R^(∘))C(O)NR^(∘))₂;—N(R^(∘))N(R^(∘))C(O)OR^(∘); —(CH₂)₀₋₄C(O)R^(∘); —C(S)R^(∘);—(CH₂)₀₋₄C(O)OR^(∘); —(CH₂)₀₋₄C(O)SR^(∘); —(CH₂)₀₋₄C(O)O SiR^(∘) ₃;—(CH₂)₀₋₄OC(O)R^(∘); —OC(O)(CH₂)₀₋₄SR—, SC(S) SR^(∘);—(CH₂)₀₋₄SC(O)R^(∘); —(CH₂)₀₋₄C(O)NR^(∘))₂; —C(S)NR^(∘))₂; —C(S)SR^(∘);—SC(S)SR^(∘), —(CH₂)₀₋₄OC(O)NR^(∘))₂; —C(O)N(OR^(∘))R^(∘);—C(O)C(O)R^(∘); —C(O)CH₂C(O)R^(∘); —C(NOR^(∘))R^(∘); —(CH₂)₀₋₄SSR^(∘);—(CH₂)₀₋₄S(O)₂R^(∘); —(CH₂)₀₋₄S(O)₂OR^(∘); —(CH₂)₀₋₄OS(O)₂R^(∘);—S(O)₂NR^(∘))₂; —S(O)(NR^(∘))R^(∘); —S(O)₂N═C(NR^(∘))₂)₂;—(CH₂)₀₋₄S(O)R^(∘); —)N(R^(∘))S(O)₂NR^(∘))₂; —N(R^(∘))S(O)₂R^(∘);—N(OR^(∘))R^(∘); —C(NH)NR^(∘))₂; —P(O)₂R^(∘); —P(O)R^(∘))₂;—OP(O)R^(∘))₂; —OP(O)(OR^(∘))₂; SiR^(∘) ₃; —(C₁₋₄ straight or branched)alkylene)O—N(R^(∘))₂; or —(C₁₋₄ straight orbranched)alkylene)C(O)O—N(R^(∘))₂.

Each R^(∘) is independently hydrogen, C₁₋₆ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, —CH₂-(5-6 membered heteroaryl ring), or a 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or,notwithstanding the definition above, two independent occurrences ofR^(∘), taken together with their intervening atom(s), form a3-12-membered saturated, partially unsaturated, or aryl mono- orbicyclic ring having 0-4 heteroatoms independently selected fromnitrogen, oxygen, or sulfur, which may be substituted by a divalentsubstituent on a saturated carbon atom of R^(∘) selected from ═O and ═S;or each R^(∘) is optionally substituted with a monovalent substituentindependently selected from halogen, —(CH₂)₀₋₂R^(•), -(haloR^(•)),—(CH₂)₀₋₂OH, —(CH₂)₀₋₂OR^(•), —(CH₂)₀₋₂CH(OR^(•))₂; —O(haloR^(•)), —CN,—N₃, —(CH₂)₀₋₂C(O)R^(•), —(CH₂)₀₋₂C(O)OH, —(CH₂)₀₋₂C(O)OR^(•),—(CH₂)₀₋₂SR^(•), —(CH₂)₀₋₂SH, —(CH₂)₀₋₂NH₂, —(CH₂)₀₋₂NHR^(•),—(CH₂)₀₋₂NR^(•) ₂, —NO₂, —SiR^(•) ₃, —OSiR^(•) ₃, —C(O)SR^(•), —(C₁₋₄straight or branched alkylene)C(O)OR^(•), or —SSR^(•).

Each R^(•) is independently selected from C₁₋₄ aliphatic, —CH₂Ph,—O(CH₂)₀₋₁Ph, or a 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur, and wherein each R^(•) is unsubstituted or wherepreceded by halo is substituted only with one or more halogens; orwherein an optional substituent on a saturated carbon is a divalentsubstituent independently selected from ═O, ═S, ═NNR^(•) ₂, ═NNHC(O)R*,═NNHC(O)OR*, ═NNHS(O)₂R*, ═NR*, ═NOR*, —O(C(R*₂))₂₋₃O—, or —S(C(R*2))₂₋₃S—, or a divalent substituent bound to vicinal substitutable carbons ofan “optionally substituted” group is —O(CR*₂)₂₋₃O—, wherein eachindependent occurrence of R* is selected from hydrogen, C₁₋₆ aliphaticor an unsubstituted 5-6-membered saturated, partially unsaturated, oraryl ring having 0-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

When R* is C₁₋₆ aliphatic, R* is optionally substituted with halogen,—R^(•), -(haloR^(•)), —OH, —OR^(•), —O(haloR^(•)), —CN, —C(O)OH,—C(O)OR^(•), —NH₂, —NHR^(•), —NR^(•) ₂, or —NO₂, wherein each R^(•) isindependently selected from C₁₋₄ aliphatic, —CH₂Ph, —O(CH₂)₀₋₁Ph, or a5-6-membered saturated, partially unsaturated, or aryl ring having 0-4heteroatoms independently selected from nitrogen, oxygen, or sulfur, andwherein each R^(•) is unsubstituted or where preceded by halo issubstituted only with one or more halogens.

An optional substituent on a substitutable nitrogen is independently—R^(†), —C(O)R^(†), —C(O)OR^(†), —C(O)C(O)R^(†), —C(O)CH₂C(O)R^(†),—S(O)₂R^(†), —S(O)₂NR^(†) ₂, —C(S)NR^(†) ₂, —C(NH)NR^(†) ₂, or—N(R^(†))S(O)₂R^(†); wherein each Rt is independently hydrogen, C₁₋₆aliphatic, unsubstituted —OPh, or an unsubstituted 5-6-memberedsaturated, partially unsaturated, or aryl ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or, twoindependent occurrences of R^(†), taken together with their interveningatom(s) form an unsubstituted 3-12-membered saturated, partiallyunsaturated, or aryl mono- or bicyclic ring having 0-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur; wherein whenR^(†) is C₁₋₆ aliphatic, R^(†) is optionally substituted with halogen,−R^(•), -(haloR^(•)), —OH, −OR^(•), —O(haloR^(•)), —CN, —C(O)OH,—C(O)OR^(•), —NH₂, —NHR^(•), —NR^(•) ₂, or —NO₂, wherein each R^(•) isindependently selected from C₁₋₄ aliphatic, —CH₂Ph, —O(CH₂)₀₋₁Ph, or a5-6-membered saturated, partially unsaturated, or aryl ring having 0-4heteroatoms independently selected from nitrogen, oxygen, or sulfur, andwherein each R^(•) is unsubstituted or where preceded by halo issubstituted only with one or more halogens.

As used herein, the term “pharmaceutically acceptable salt” refers tothose salts which are, within the scope of sound medical judgment,suitable for use in contact with the tissues of humans and lower animalswithout undue toxicity, irritation, allergic response and the like, andare commensurate with a reasonable benefit/risk ratio. Pharmaceuticallyacceptable salts are well known in the art. For example, S. M. Berge etal., describe pharmaceutically acceptable salts in detail in J.Pharmaceutical Sciences, 1977, 66, 1-19, incorporated herein byreference. Pharmaceutically acceptable salts of the compounds of thisinvention include those derived from suitable inorganic and organicacids and bases. Examples of pharmaceutically acceptable, nontoxic acidaddition salts are salts of an amino group formed with inorganic acidssuch as hydrochloric acid, hydrobromic acid, phosphoric acid, sulfuricacid and perchloric acid or with organic acids such as acetic acid,oxalic acid, maleic acid, tartaric acid, citric acid, succinic acid ormalonic acid or by using other methods used in the art such as ionexchange. Other pharmaceutically acceptable salts include adipate,alginate, ascorbate, aspartate, benzenesulfonate, benzoate, bisulfate,borate, butyrate, camphorate, camphorsulfonate, citrate,cyclopentanepropionate, digluconate, dodecylsulfate, ethanesulfonate,formate, fumarate, glucoheptonate, glycerophosphate, gluconate,hemisulfate, heptanoate, hexanoate, hydroiodide,2-hydroxy-ethanesulfonate, lactobionate, lactate, laurate, laurylsulfate, malate, maleate, malonate, methanesulfonate,2-naphthalenesulfonate, nicotinate, nitrate, oleate, oxalate, palmitate,pamoate, pectinate, persulfate, 3-phenylpropionate, phosphate, pivalate,propionate, stearate, succinate, sulfate, tartrate, thiocyanate,p-toluenesulfonate, undecanoate, valerate salts, and the like.

Salts derived from appropriate bases include alkali metal, alkalineearth metal, ammonium and N⁺(C₁₋₄ alkyl)₄ salts. Representative alkalior alkaline earth metal salts include sodium, lithium, potassium,calcium, magnesium, and the like. Further pharmaceutically acceptablesalts include, when appropriate, nontoxic ammonium, quaternary ammonium,and amine cations formed using counterions such as halide, hydroxide,carboxylate, sulfate, phosphate, nitrate, loweralkyl sulfonate and arylsulfonate.

Unless otherwise stated, structures depicted herein are also meant toinclude all isomeric (e.g., enantiomeric, diastereomeric, and geometric(or conformational)) forms of the structure; for example, the R and Sconfigurations for each asymmetric center, Z and E double bond isomers,and Z and E conformational isomers. Therefore, single stereochemicalisomers as well as enantiomeric, diastereomeric, and geometric (orconformational) mixtures of the present compounds are within the scopeof the invention. Unless otherwise stated, all tautomeric forms of thecompounds of the invention are within the scope of the invention.Additionally, unless otherwise stated, structures depicted herein arealso meant to include compounds that differ only in the presence of oneor more isotopically enriched atoms. For example, compounds having thepresent structures including the replacement of hydrogen by deuterium ortritium, or the replacement of a carbon by a ¹³C- or ¹⁴C-enriched carbonare within the scope of this invention. Such compounds are useful, forexample, as analytical tools, as probes in biological assays, or astherapeutic agents in accordance with the present invention. In certainembodiments, a warhead moiety, R¹, of a provided compound comprises oneor more deuterium atoms.

As used herein, the term “inhibitor” is defined as a compound that bindsto and/or inhibits CXCR4 with measurable affinity. In certainembodiments, an inhibitor has an IC₅₀ and/or binding constant of lessthan about 100 μM, less than about 50 μM, less than about 1 μM, lessthan about 500 nM, less than about 100 nM, less than about 10 nM, orless than about 1 nM.

The terms “measurable affinity” and “measurably inhibit,” as usedherein, means a measurable change in CXCR4 activity between a samplecomprising a compound of the present invention, or composition thereof,and CXCR4, and an equivalent sample comprising CXCR4, in the absence ofsaid compound, or composition thereof.

3. Description of Exemplary Embodiments

In one aspect, the present invention provides a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein:

-   Ring A is a 3-8 membered saturated or partially unsaturated    monocyclic carbocyclic ring, phenyl, an 8-10 membered bicyclic    aromatic carbocyclic ring, a 4-8 membered saturated or partially    unsaturated monocyclic heterocyclic ring having 1-2 heteroatoms    independently selected from nitrogen, oxygen, or sulfur, a 5-6    membered monocyclic heteroaromatic ring having 1-4 heteroatoms    independently selected from nitrogen, oxygen, or sulfur, or an 8-10    membered bicyclic heteroaromatic ring having 1-5 heteroatoms    independently selected from nitrogen, oxygen, or sulfur;-   each R¹ is independently —R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃,    —SR, or -L¹-R⁶; each R is independently hydrogen or an optionally    substituted group selected from C₁₋₆ aliphatic, a 3-8 membered    saturated or partially unsaturated monocyclic carbocyclic ring,    phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 4-8    membered saturated or partially unsaturated monocyclic heterocyclic    ring having 1-2 heteroatoms independently selected from nitrogen,    oxygen, or sulfur, a 5-6 membered monocyclic heteroaromatic ring    having 1-4 heteroatoms independently selected from nitrogen, oxygen,    or sulfur, or an 8-10 membered bicyclic heteroaromatic ring having    1-5 heteroatoms independently selected from nitrogen, oxygen, or    sulfur;-   each L¹ and L² is independently a covalent bond or a C₁₋₈ bivalent    straight or branched hydrocarbon chain wherein 1, 2, or 3 methylene    units of the chain are independently and optionally replaced with    —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—, —(R)NC(O)—, —OC(O)N(R)—,    —(R)NC(O)O—, —N(R)C(O)N(R)—, —S—, —SO—, —SO₂N(R)—, —(R)NSO₂—,    —C(S)—, —C(S)O—, —OC(S)—, —C(S)N(R)—, —(R)NC(S)—, —(R)NC(S)N(R)—, or    -Cy-;-   each -Cy- is independently a bivalent optionally substituted 3-8    membered saturated or partially unsaturated monocyclic carbocyclic    ring, optionally substituted phenylene, an optionally substituted    4-8 membered saturated or partially unsaturated monocyclic    heterocyclic ring having 1-3 heteroatoms independently selected from    nitrogen, oxygen, or sulfur, an optionally substituted 5-6 membered    monocyclic heteroaromatic ring having 1-4 heteroatoms independently    selected from nitrogen, oxygen, or sulfur, an optionally substituted    8-10 membered bicyclic or bridged bicyclic saturated or partially    unsaturated heterocyclic ring having 1-5 heteroatoms independently    selected from nitrogen, oxygen, or sulfur, or an optionally    substituted 8-10 membered bicyclic or bridged bicyclic    heteroaromatic ring having 1-5 heteroatoms independently selected    from nitrogen, oxygen, or sulfur;-   R² is hydrogen, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃, —SR, -L²-R⁶,    or optionally substituted C₁₋₈ aliphatic;-   R³ is hydrogen, optionally substituted C₁₋₆ aliphatic, or -L³-R⁶;-   L³ is a C₁₋₆ bivalent straight or branched hydrocarbon chain wherein    1, 2, or 3 methylene units of the chain are independently and    optionally replaced with —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—,    —C(O)N(R)—, —(R)NC(O)—, —S—, —SO—, —C(S)—, or -Cy-;-   each R⁴ is independently hydrogen, deuterium, halogen, —CN, —OR⁶, or    C₁₋₄ alkyl, or two R⁴ groups on the same carbon are optionally taken    together to form ═NR⁶, ═NOR⁶, ═O, or ═S;-   each R⁵ is independently R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃,    —SR, or -L¹-R⁶, or two R⁵ groups on the same saturated carbon atom    are optionally taken together to form ═NR, ═NOR, ═O, ═S, or a    spirocyclic 3-6 membered carbocyclic ring;-   each R⁶ is independently hydrogen or C₁₋₆ alkyl optionally    substituted with 1, 2, 3, 4, 5, or 6 deuterium or halogen atoms;-   m is 0, 1, 2, 3, or 4;-   n is 0, 1, 2, 3, or 4; and-   p is 0, 1, 2, 3, or 4.

As defined generally above, Ring A is a 3-8 membered saturated orpartially unsaturated monocyclic carbocyclic ring, phenyl, an 8-10membered bicyclic aromatic carbocyclic ring, a 4-8 membered saturated orpartially unsaturated monocyclic heterocyclic ring having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur, a5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or an 8-10membered bicyclic heteroaromatic ring having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur.

In some embodiments, Ring A is a 3-8 membered saturated or partiallyunsaturated monocyclic carbocyclic ring. In some embodiments, Ring A isphenyl. In some embodiments, Ring A is an 8-10 membered bicyclicaromatic carbocyclic ring. In some embodiments, Ring A is a 4-8 memberedsaturated or partially unsaturated monocyclic heterocyclic ring having1-2 heteroatoms independently selected from nitrogen, oxygen, or sulfur.In some embodiments, Ring A is a 5-6 membered monocyclic heteroaromaticring having 1-4 heteroatoms independently selected from nitrogen,oxygen, or sulfur. In some embodiments, Ring A is an 8-10 memberedbicyclic heteroaromatic ring having 1-5 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur.

In some embodiments, Ring A is a 5-6 membered monocyclic heteroaromaticring having 1-2 heteroatoms independently selected from nitrogen,oxygen, or sulfur.

In some embodiments, Ring A is selected from:

In some embodiments, Ring A is selected from

In some embodiments, Ring A is selected from

In some embodiments, Ring A is

In some embodiments, Ring A is not

In some embodiments, Ring A is not imidazo[1,2-a]pyridine.

In some embodiments, Ring A is selected from those depicted in Table 1,below.

As defined generally above, each R¹ is independently R, halogen, —CN,—OR, —N(R)₂, —NO2, —N₃, —SR, or -L¹-R⁶.

In some embodiments, R¹ is R. In some embodiments, R¹ is halogen. Insome embodiments, R¹ is —CN. In some embodiments, R¹ is —OR. In someembodiments, R¹ is —N(R)₂. In some embodiments, R¹ is —NO₂. In someembodiments, R¹ is —N₃. In some embodiments, le is —SR. In someembodiments, R¹ is -L¹-R⁶.

In some embodiments, R¹ is hydrogen. In some embodiments, R¹ is anoptionally substituted C₁₋₆ aliphatic group. In some embodiments, R¹ isan optionally substituted 3-8 membered saturated or partiallyunsaturated monocyclic carbocyclic ring. In some embodiments, R¹ is anoptionally substituted phenyl. In some embodiments, R¹ is an optionallysubstituted 8-10 membered bicyclic aromatic carbocyclic ring. In someembodiments, R¹ is an optionally substituted 4-8 membered saturated orpartially unsaturated monocyclic heterocyclic ring having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur. Insome embodiments, R¹ is an optionally substituted 5-6 memberedmonocyclic heteroaromatic ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur. In some embodiments, R¹ is anoptionally substituted 8-10 membered bicyclic heteroaromatic ring having1-5 heteroatoms independently selected from nitrogen, oxygen, or sulfur.

In some embodiments, R¹ is selected from R, halogen, —CN, —OR, —N(R)₂,—SR, C₁₋₆ aliphatic, or -L¹-R⁶, wherein L¹ is a C₁₋₆ bivalent straightor branched hydrocarbon chain wherein 1, 2, or 3 methylene units of thechain are independently and optionally replaced with —O—, —C(O)—,—N(R)—, —S—, —SO—, —SO₂—, —C(S)—, or -Cy-; wherein the C₁₋₆ aliphaticgroup is optionally substituted with 1, 2, or 3 groups independentlyselected from halogen, —CN, —N(R)₂, —NO₂, —N₃, ═NR, ═NOR, ═O, ═S, —OR,—SR, —SO₂R, —S(O)R, —R, -Cy-R, —C(O)R, —C(O)OR, —OC(O)R, —C(O)N(R)₂,—(R)NC(O)R, —OC(O)N(R)₂, —(R)NC(O)OR, —N(R)C(O)N(R)₂, —SO₂N(R)₂,—(R)NSO₂R, —C(S)R, or —C(S)OR; and each R is independently hydrogen,—CH₂-phenyl, phenyl, C₁₋₆ alkyl, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, —CH₂F, —CHF₂, —CF₃, —CH₂CHF₂, or —CH₂CF₃; or each R isindependently hydrogen or methyl; or R is hydrogen.

In some embodiments, R¹ is selected from hydrogen, halogen, C₁₋₆ alkyl(optionally substituted with 1, 2, or 3 halogens), —CN, —N(R)₂, —OR,—SR, —S(O)R⁶, —SO₂R⁶, —SO₂NHR⁶,

and each R is independently hydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃,—CH₂CHF₂, or —CH₂CF₃; or each R is independently hydrogen or methyl; orR is hydrogen.

In some embodiments, R¹ is selected from hydrogen, halogen, C₁₋₆ alkyl,—CN, —N(R)₂, —OR, —SR,

and each R is independently hydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃,—CH₂CHF₂, or —CH₂CF₃; or each R is independently hydrogen or methyl; orR is hydrogen.

In some embodiments, R¹ is selected from those depicted in Table 1,below.

As defined generally above, each L¹ and L² is independently a covalentbond or a C₁₋₈ bivalent straight or branched hydrocarbon chain wherein1, 2, or 3 methylene units of the chain are independently and optionallyreplaced with —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—, —C(O)N(R)—,—(R)NC(O)—, —OC(O)N(R)—, —(R)NC(O)O—, —N(R)C(O)N(R)—, —S—, —SO—, —SO₂—,—SO₂N(R)—, —(R)NSO₂—, —C(S)—, —C(S)O—, —OC(S)—, —C(S)N(R)—, —(R)NC(S)—,—(R)NC(S)N(R)—, or -Cy-.

In some embodiments, L¹ is a covalent bond. In some embodiments, L¹ is aC₁₋₈ bivalent straight or branched hydrocarbon chain. In someembodiments, L¹ is a C18 bivalent straight or branched hydrocarbon chainwherein 1, 2, or 3 methylene units of the chain are independently andoptionally replaced with —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—,—(R)NC(O)—, —OC(O)N(R)—, —(R)NC(O)O—, —N(R)C(O)N(R)—, —S—, —SO—,—SO₂N(R)—, —(R)NSO₂—, —C(S)—, —C(S)O—, —OC(S)—, —C(S)N(R)—, —(R)NC(S)—,—(R)NC(S)N(R)—, or -Cy-.

In some embodiments, L¹ is a C₁₋₆ bivalent straight or branchedhydrocarbon chain wherein 1, 2, or 3 methylene units of the chain areindependently and optionally replaced with —O—, —C(O)—, —N(R)—, —S—,—SO—, —SO₂N(R)—, —(R)NSO₂—, —C(S)—, or -Cy-, and each R is independentlyhydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃, —CH₂CHF₂, or —CH₂CF₃; oreach R is independently hydrogen or methyl; or R is hydrogen.

In some embodiments, L¹ is selected from those depicted in Table 1,below.

In some embodiments, L² is a covalent bond. In some embodiments, L² is aC₁₋₈ bivalent straight or branched hydrocarbon chain. In someembodiments, L² is a C18 bivalent straight or branched hydrocarbon chainwherein 1, 2, or 3 methylene units of the chain are independently andoptionally replaced with —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—,—(R)NC(O)—, —OC(O)N(R)—, —(R)NC(O)O—, —N(R)C(O)N(R)—, —S—, —SO—,—SO₂N(R)—, —(R)NSO₂—, —C(S)—, —C(S)O—, —OC(S)—, —C(S)N(R)—, —(R)NC(S)—,—(R)NC(S)N(R)—, or -Cy-.

In some embodiments, L² is a C₁₋₆ bivalent straight or branchedhydrocarbon chain wherein 1, 2, or 3 methylene units of the chain areindependently and optionally replaced with —O—, —C(O)—, —N(R)—, —S—,—SO—, —SO₂N(R)—, —(R)NSO₂—, —C(S)—, or -Cy-, and each R is independentlyhydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl, cyclopropyl, cyclobutyl,cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃, —CH₂CHF₂, or —CH₂CF₃; oreach R is independently hydrogen or methyl; or R is hydrogen.

In some embodiments, L² is selected from those depicted in Table 1,below.

As defined generally above, each -Cy- is independently a bivalentoptionally substituted 3-8 membered saturated or partially unsaturatedmonocyclic carbocyclic ring, optionally substituted phenylene, anoptionally substituted 4-8 membered saturated or partially unsaturatedmonocyclic heterocyclic ring having 1-3 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur, an optionally substituted 5-6membered monocyclic heteroaromatic ring having 1-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, an optionallysubstituted 8-10 membered bicyclic or bridged bicyclic saturated orpartially unsaturated heterocyclic ring having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or anoptionally substituted 8-10 membered bicyclic or bridged bicyclicheteroaromatic ring having 1-5 heteroatoms independently selected fromnitrogen, oxygen, or sulfur.

In some embodiments, -Cy- is a bivalent optionally substituted 3-8membered saturated or partially unsaturated monocyclic carbocyclic ring.In some embodiments, -Cy- is an optionally substituted phenylene. Insome embodiments, -Cy- is an optionally substituted 4-8 memberedsaturated or partially unsaturated monocyclic heterocyclic ring having1-3 heteroatoms independently selected from nitrogen, oxygen, or sulfur.In some embodiments, -Cy- is an optionally substituted 5-6 memberedmonocyclic heteroaromatic ring having 1-4 heteroatoms independentlyselected from nitrogen, oxygen, or sulfur. In some embodiments, -Cy- isan optionally substituted 8-10 membered bicyclic or bridged bicyclicsaturated or partially unsaturated heterocyclic ring having 1-5heteroatoms independently selected from nitrogen, oxygen, or sulfur. Insome embodiments, -Cy- is an optionally substituted 8-10 memberedbicyclic or bridged bicyclic heteroaromatic ring having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur.

In some embodiments, -Cy- is

In some embodiments, -Cy- is selected from those depicted in Table 1,below.

As defined generally above, R² is hydrogen, halogen, —CN, —OR, —N(R)₂,—NO₂, —N₃, —SR, -L²-R⁶, or optionally substituted C₁₋₈ aliphatic.

In some embodiments, R² is hydrogen. In some embodiments, R² is halogen.In some embodiments, R² is —CN. In some embodiments, R² is —OR. In someembodiments, R² is —N(R)₂. In some embodiments, R² is —NO₂. In someembodiments, R² is —N₃. In some embodiments, R² is —SR. In someembodiments, R² is -L²-R⁶. In some embodiments, R² is optionallysubstituted C₁₋₈ aliphatic.

In some embodiments, R² is hydrogen, halogen, —CN, —OR, —N(R)₂, —SR,optionally substituted C₁₋₆ aliphatic, or -L²-R⁶, wherein L² is a C₁₋₆bivalent straight or branched hydrocarbon chain wherein 1, 2, or 3methylene units of the chain are independently and optionally replacedwith —O—, —C(O)—, —N(R)—, —S—, —SO—, —SO₂—, —C(S)—, or -Cy-; wherein theC₁₋₆ aliphatic group is optionally substituted with 1, 2, or 3 groupsindependently selected from halogen, —CN, —N(R)₂, —NO₂, —N₃, ═NR, ═NOR,═O, ═S, —OR, —SR, —SO₂R, —S(O)R, —R, -Cy-R, —C(O)R, —C(O)OR, —OC(O)R,—C(O)N(R)₂, —(R)NC(O)R, —OC(O)N(R)₂, —(R)NC(O)OR, —N(R)C(O)N(R)₂,—SO₂N(R)₂, —(R)NSO₂R, —C(S)R, or —C(S)OR; wherein each R isindependently hydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃, —CH₂CHF₂, or—CH₂CF₃; or each R is independently hydrogen or methyl; or R ishydrogen.

In some embodiments, R² is selected from hydrogen, halogen, —CN, —OR,—N(R)₂, C₁₋₆ alkyl (optionally substituted with 1, 2, or 3 deuterium orhalogen atoms), C₂₋₆ alkynyl, —S(O)R⁶, —SO₂R⁶, —SO₂NHR⁶,—(CH₂)₁₋₆—N(R)R⁶, —(CH₂)₁₋₆—OR⁶, or —(CH₂)₀₋₆-Cy-R⁶. In someembodiments, R² is selected from hydrogen, halogen, —OR, —N(R)₂,—S(O)R⁶, —SO₂R⁶, —SO₂NHR⁶, —(CH₂)₁₋₆—N(R)R⁶, —(CH₂)₁₋₆—OR⁶,

and each R is independently hydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃,—CH₂CHF₂, or —CH₂CF₃; or each R is independently hydrogen or methyl; orR is hydrogen.

In some embodiments, R² is C₂₋₄ alkynyl, —NH₂, F, Cl, Br, or I. In someembodiments, R² is hydrogen, Cl, —NH₂, or ethynyl. In some embodiments,R² is Cl.

In some embodiments, R² is selected from those depicted in Table 1,below.

As defined generally above, R³ is hydrogen, optionally substituted C₁₋₆aliphatic, or -L³-R⁶.

In some embodiments, R³ is hydrogen. In some embodiments, R³ isoptionally substituted C₁₋₆ aliphatic. In some embodiments, R³ is-L³-R⁶.

In some embodiments, R³ is selected from hydrogen or C₁₋₆ alkyloptionally substituted with 1, 2, or 3 groups independently selectedfrom deuterium, halogen, —CN, —N(R)₂, —NO2, —N₃, ═NR, ═NOR, ═O, ═S, —OR,—SR, —SO₂R, —S(O)R, —R, -Cy-R, —C(O)R, —C(O)OR, —OC(O)R, —C(O)N(R)₂,—(R)NC(O)R, —OC(O)N(R)₂, —(R)NC(O)OR, —N(R)C(O)N(R)₂, —SO₂N(R)₂,—(R)NSO₂R, —C(S)R, or —C(S)OR. In some embodiments, R³ is selected fromhydrogen or C₁₋₆ alkyl (optionally substituted with 1, 2, or 3 deuteriumor halogen atoms), —(CH₂)₁₋₆—CN, —(CH₂)₁₋₆—N(R)(R⁶), —(CH₂)₁₋₆—OR⁶, or—(CH₂)₀₋₆-Cy-R⁶. In some embodiments, R³ is selected from hydrogen, C₁₋₆alkyl (optionally substituted with 1, 2, or 3 deuterium or halogenatoms), —(CH₂)₁₆—CN, —(CH₂)₁₆—N(R)(R⁶), —(CH₂)₁₆—OR⁶,

and each R is independently hydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃,—CH₂CHF₂, or —CH₂CF₃; or each R is independently hydrogen or methyl; orR is hydrogen.

In some embodiments, R³ is hydrogen or C₁₋₆ alkyl optionally substitutedwith 1, 2, or 3 deuterium or halogen atoms, phenyl, pyridyl, —CN,—N(R)₂, or —OR, wherein each R is independently hydrogen, —CH₂-phenyl,phenyl, C₁₋₆ alkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,—CH₂F, —CHF₂, —CF₃, —CH₂CHF₂, or —CH₂CF₃; or each R is independentlyhydrogen or methyl; or R is hydrogen. In some embodiments, R³ is C₁₋₄alkyl optionally substituted with

pyridyl, —N(R)₂, —CN, or 1, 2, or 3 deuterium or halogen atoms, whereinR is hydrogen or C₁₋₃ alkyl. In some embodiments, R³ is methyl, ethyl,—CD₃, or —CH₂CF₃. In some embodiments, R³ is methyl.

In some embodiments, R³ is selected from those depicted in Table 1,below.

As defined generally above, L³ is a C₁₋₆ bivalent straight or branchedhydrocarbon chain wherein 1, 2, or 3 methylene units of the chain areindependently and optionally replaced with —O—, —C(O)—, —C(O)O—,—OC(O)—, —N(R)—, —C(O)N(R)—, —(R)NC(O)—, —S—, —SO—, —SO₂—, —C(S)—, or-Cy-.

In some embodiments, L³ is a C₁₋₆ bivalent straight or branchedhydrocarbon chain. In some embodiments, L³ is a C₁₋₆ bivalent straightor branched hydrocarbon chain wherein 1, 2, or 3 methylene units of thechain are independently and optionally replaced with —O—, —C(O)—,—C(O)O—, —OC(O)—, —N(R)—, —C(O)N(R)—, —(R)NC(O)—, —S—, —SO—, —SO₂—,—C(S)—, or -Cy-.

In some embodiments, L³ is selected from those depicted in Table 1,below.

As defined generally above, each R⁴ is independently hydrogen,deuterium, halogen, —CN, —OR⁶, or C₁₋₄ alkyl, or two R⁴ groups on thesame carbon are optionally taken together to form ═NR⁶, ═NOR⁶, ═O, or═S.

In some embodiments, R⁴ is hydrogen. In some embodiments, R⁴ isdeuterium. In some embodiments, R⁴ is halogen. In some embodiments, R⁴is —CN. In some embodiments, R⁴ is —OR⁶. In some embodiments, R⁴ is C₁₋₄alkyl. In some embodiments, two R⁴ groups on the same carbon areoptionally taken together to form ═NR⁶, ═NOR⁶, ═O, or ═S.

In some embodiments, R⁴ is hydrogen, deuterium, halogen, —CN, C₁₋₂alkyl, or two R⁴ groups on the same carbon are taken together to form ═Oor ═S.

In some embodiments, R⁴ is selected from those depicted in Table 1,below.

As defined generally above, each R⁵ is independently R, halogen, —CN,—OR, —N(R)₂, —NO2, —N₃, —SR, or -L¹-R⁶, or two R⁵ groups on the samesaturated carbon atom are optionally taken together to form ═NR, ═NOR,═O, ═S, or a spirocyclic 3-6 membered carbocyclic ring.

In some embodiments, R⁵ is R. In some embodiments, R⁵ is halogen. Insome embodiments, R⁵ is —CN. In some embodiments, R⁵ is —OR. In someembodiments, R⁵ is —N(R)₂.

In some embodiments, R⁵ is —NO₂. In some embodiments, R⁵ is —N₃. In someembodiments, R⁵ is —SR. In some embodiments, R⁵ is —O—R⁶. In someembodiments, two R⁵ groups on the same saturated carbon atom are takentogether to form ═NR, ═NOR, ═O, ═S, or a spirocyclic 3-6 memberedcarbocyclic ring.

In some embodiments, R⁵ is hydrogen. In some embodiments, R⁵ is anoptionally substituted C₁₋₆ aliphatic group. In some embodiments, R⁵ isa C₁₋₆ alkyl group optionally substituted with 1, 2, 3, or 4 deuteriumor halogen atoms. In some embodiments, R⁵ is an optionally substituted3-8 membered saturated or partially unsaturated monocyclic carbocyclicring. In some embodiments, R⁵ is an optionally substituted phenyl. Insome embodiments, R⁵ is an optionally substituted 8-10 membered bicyclicaromatic carbocyclic ring. In some embodiments, R⁵ is an optionallysubstituted 4-8 membered saturated or partially unsaturated monocyclicheterocyclic ring having 1-2 heteroatoms independently selected fromnitrogen, oxygen, or sulfur. In some embodiments, R⁵ is an optionallysubstituted 5-6 membered monocyclic heteroaromatic ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur. Insome embodiments, R⁵ is an optionally substituted 8-10 membered bicyclicheteroaromatic ring having 1-5 heteroatoms independently selected fromnitrogen, oxygen, or sulfur.

In some embodiments, R⁵ is hydrogen, C₁₋₆ alkyl, halogen, —CN, —CF₃,-CD3, cyclopropyl, ethynyl, —OCH₃, —OCF₃, or

In some embodiments, R⁵ is methyl.

In some embodiments, R⁵ is selected from those depicted in Table 1,below.

As defined generally above, each R⁶ is independently hydrogen or C₁₋₆alkyl optionally substituted with 1, 2, 3, 4, 5, or 6 deuterium orhalogen atoms.

In some embodiments, R⁶ is hydrogen. In some embodiments, R⁶ is C₁₋₆alkyl optionally substituted with 1, 2, 3, 4, 5, or 6 deuterium orhalogen atoms.

In some embodiments, R⁶ is C₁₋₃ alkyl optionally substituted with 1, 2,or 3 deuterium or halogen atoms.

In some embodiments, R⁶ is selected from those depicted in Table 1,below.

As defined generally above, m is 0, 1, 2, 3, or 4. In some embodiments,m is 0. In some embodiments, m is 1. In some embodiments, m is 2. Insome embodiments, m is 3. In some embodiments, m is 4. In someembodiments, m is 0, 1, 2, or 3. In some embodiments, m is 0, 1, or 2.In some embodiments, m is 1, 2, or 3.

As defined generally above, n is 0, 1, 2, 3, or 4. In some embodiments,n is 0. In some embodiments, n is 1. In some embodiments, n is 2. Insome embodiments, n is 3. In some embodiments, n is 4. In someembodiments, n is 0, 1, 2, or 3. In some embodiments, n is 0, 1, or 2.In some embodiments, n is 1, 2, or 3.

As defined generally above, p is 0, 1, 2, 3, or 4. In some embodiments,p is 0. In some embodiments, p is 1. In some embodiments, p is 2. Insome embodiments, p is 3. In some embodiments, p is 4. In someembodiments, p is 0, 1, 2, or 3. In some embodiments, p is 0, 1, or 2.In some embodiments, p is 1, 2, or 3.

In some embodiments, the present invention provides a compound ofFormulae II-a or II-b:

or a pharmaceutically acceptable salt thereof, wherein each of Ring A,R, R¹, R², R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, m, n, and p is as definedabove and described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound ofFormula III:

or a pharmaceutically acceptable salt thereof, wherein each of Ring A,R, R¹, R², R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, n, and p is as definedabove and described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound ofFormula IV:

or a pharmaceutically acceptable salt thereof, wherein each of Ring A,R, R¹, R², R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, m, and p is as definedabove and described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound ofFormula V:

or a pharmaceutically acceptable salt thereof, wherein each of Ring A,R, R¹, R², R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, m, and p is as definedabove and described in embodiments herein, both singly and incombination.

In some embodiments, the present invention provides a compound ofFormula VI:

or a pharmaceutically acceptable salt thereof, wherein each of R, R¹,R², R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, m, n, and p is as defined aboveand described in embodiments herein, both singly and in combination.

In some embodiments, the present invention provides a compound ofFormula VII:

or a pharmaceutically acceptable salt thereof, wherein each of R, R¹,R², R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, m, n, and p is as defined aboveand described in embodiments herein, both singly and in combination.

In some embodiments, the present invention provides a compound ofFormulae VIII-a or VIII-b:

or a pharmaceutically acceptable salt thereof, wherein each of R, R²,R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, m, n, and p is as defined above anddescribed in embodiments herein, both singly and in combination.

In some embodiments, the present invention provides a compound ofFormula IX:

or a pharmaceutically acceptable salt thereof, wherein each of R, R²,R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, and n is as defined above anddescribed in embodiments herein, both singly and in combination.

In some embodiments, the present invention provides a compound ofFormulae X-a, X-b, X-c, X-d, or X-e:

or a pharmaceutically acceptable salt thereof, wherein each of R, R¹,R², R³, R⁴, R⁵, R⁶, L¹, L², L³, -Cy-, and n is as defined above anddescribed in embodiments herein, both singly and in combination.

In some embodiments, the present invention provides a compound ofFormula XI:

or a pharmaceutically acceptable salt thereof, wherein each of R, R¹,R², R³, R⁵, R⁶, L¹, L², L³, -Cy-, and p is as defined above anddescribed in embodiments herein, both singly and in combination.

In some embodiments, the present invention provides a compound ofFormulae XII-a or XII-b:

or a pharmaceutically acceptable salt thereof, wherein each of R, R¹,R², R³, R⁵, R⁶, L¹, L², L³, -Cy-, and p is as defined above anddescribed in embodiments herein, both singly and in combination.

In some embodiments, the present invention provides a compound ofFormulae XIII-a or XIII-b:

or a pharmaceutically acceptable salt thereof, wherein each of R, R²,R³, R⁶, L², L³, and -Cy- is as defined above and described inembodiments herein, both singly and in combination. In some embodimentsof Formulae XIII-a or XIII-b, -Cy- is an optionally substituted 4-8membered saturated or partially unsaturated monocyclic heterocyclic ringhaving 1-3 heteroatoms independently selected from nitrogen, oxygen, orsulfur. In some embodiments, -Cy- is a 5- or 6-membered saturated orpartially unsaturated monocyclic heterocyclic ring having 2 nitrogenatoms.

In some embodiments, the present invention provides a compound ofFormulae XIV-a, XIV-b, or XIV-c:

or a pharmaceutically acceptable salt thereof, wherein each of R, R²,R³, R⁵, R⁶, L², L³, and -Cy- is as defined above and described inembodiments herein, both singly and in combination. In some embodimentsof Formulae XIV-a, XIV-b, and XIV-c, R² is selected from hydrogen orhalogen. In some embodiments, R² is halogen. In some embodiments, R² isCl or Br. In some embodiments, R² is Cl. In some embodiments, R² is C₂₋₄alkynyl, —NH₂, F, Cl, Br, or I. In some embodiments, R² is hydrogen, Cl,—NH₂, or ethynyl.

Exemplary compounds of the invention are set forth in Table 1, below.

TABLE 1 Exemplary Compounds

I-1

I-2

I-3

I-4

I-5

I-6

I-7

I-8

I-9

I-10

I-11

I-12

I-13

I-14

I-15

I-16

I-17

I-18

I-19

I-20

I-21

I-22

I-23

I-24

I-25

I-26

I-27

I-28

I-29

I-30

I-31

I-32

I-33

I-34

I-35

I-36

I-37

I-38

I-39

I-40

I-41

I-42

I-43

I-44

I-45

I-46

I-47

I-48

I-49

I-50

I-51

I-52

I-53

I-54

I-55

I-56

I-57

I-58

I-59

I-60

I-61

I-62

I-63

I-64

I-65

I-66

I-67

I-68

I-69

I-70

I-71

I-72

I-73

I-74

I-75

I-76

I-77

I-78

I-79

I-80

I-81

I-82

I-83

I-84

I-85

I-86

I-87

I-88

I-89

I-90

I-91

I-92

I-93

I-94

I-95

I-96

I-97

I-98

I-99

I-100

I-101

I-102

I-103

I-104

I-105

I-106

I-107

I-108

I-109

I-110

I-111

I-112

I-113

I-114

I-115

I-116

I-117

I-118

I-119

I-120

I-121

I-122

I-123

I-124

I-125

I-126

I-127

I-128

I-129

I-130

I-131

I-132

I-133

I-134

I-135

I-136

I-137

I-138

I-139

I-140

I-141

I-142

I-143

I-144

I-145

I-146

I-147

I-148

I-149

I-150

I-151

I-152

I-153

I-154

I-155

I-156

I-157

I-158

I-159

I-160

I-161

I-162

I-163

I-164

I-165

I-166

I-167

I-168

I-169

I-170

I-171

I-172

I-173

I-174

I-175

I-176

I-177

I-178

I-179

I-180

I-181

I-182

I-183

I-184

I-185

I-186

I-187

I-188

I-189

I-190

I-191

I-192

I-193

I-194

I-195

I-196

I-197

I-198

I-199

I-200

I-201

I-202

I-203

I-204

I-205

I-206

I-207

I-208

I-209

I-210

I-211

I-212

I-213

I-214

I-215

I-216

I-217

I-218

I-219

I-220

I-221

I-222

I-223

I-224

I-225

I-226

I-227

I-228

I-229 Note: Stereochemistry arbitrarily assigned for I-188 and I-189;each compound was isolated in stereochemically enriched form; compoundI-149 corresponds to their racemate.

In some embodiments, the present invention provides a compound set forthin Table 1, above, or a pharmaceutically acceptable salt thereof.

4. General Methods of Providing the Present Compounds

The compounds of this invention may be prepared or isolated in generalby synthetic and/or semi-synthetic methods known to those skilled in theart for analogous compounds and by methods described in detail in theExamples, herein.

In the Schemes below, where a particular protecting group (“PG”),leaving group (“LG”), or transformation condition is depicted, one ofordinary skill in the art will appreciate that other protecting groups,leaving groups, and transformation conditions are also suitable and arecontemplated. Such groups and transformations are described in detail inMarch's Advanced Organic Chemistry: Reactions, Mechanisms, andStructure, M. B. Smith and J. March, 5^(th) Edition, John Wiley & Sons,2001, Comprehensive Organic Transformations, R. C. Larock, 2^(nd)Edition, John Wiley & Sons, 1999, and Protecting Groups in OrganicSynthesis, T. W. Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley &Sons, 1999, the entirety of each of which is hereby incorporated hereinby reference.

As used herein, the phrase “leaving group” (LG) includes, but is notlimited to, halogens (e.g. fluoride, chloride, bromide, iodide),sulfonates (e.g. mesylate, tosylate, benzenesulfonate, brosylate,nosylate, triflate), diazonium, and the like.

As used herein, the phrase “oxygen protecting group” includes, forexample, carbonyl protecting groups, hydroxyl protecting groups, etc.Hydroxyl protecting groups are well known in the art and include thosedescribed in detail in Protecting Groups in Organic Synthesis, T. W.Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley & Sons, 1999, andPhilip Kocienski, in “Protecting Groups”, Georg Thieme Verlag Stuttgart,New York, 1994, the entireties of which is incorporated herein byreference. Examples of suitable hydroxyl protecting groups include, butare not limited to, esters, allyl ethers, ethers, silyl ethers, alkylethers, arylalkyl ethers, and alkoxyalkyl ethers. Examples of suchesters include formates, acetates, carbonates, and sulfonates. Specificexamples include formate, benzoyl formate, chloroacetate,trifluoroacetate, methoxyacetate, triphenylmethoxyacetate,p-chlorophenoxyacetate, 3-phenylpropionate, 4-oxopentanoate,4,4-(ethylenedithio)pentanoate, pivaloate (trimethylacetyl), crotonate,4-methoxy-crotonate, benzoate, p-benzylbenzoate,2,4,6-trimethylbenzoate, carbonates such as methyl, 9-fluorenylmethyl,ethyl, 2,2,2-trichloroethyl, 2-(trimethylsilyl)ethyl,2-(phenylsulfonyl)ethyl, vinyl, allyl, and p-nitrobenzyl. Examples ofsuch silyl ethers include trim ethyl silyl, tri ethyl silyl, t-butyl dimethyl silyl, t-butyl di phenyl silyl, trii sopropyl silyl, and othertrialkylsilyl ethers. Alkyl ethers include methyl, benzyl,p-methoxybenzyl, 3,4-dimethoxybenzyl, trityl, t-butyl, allyl, andallyloxycarbonyl ethers or derivatives. Alkoxyalkyl ethers includeacetals such as methoxymethyl, methylthiomethyl,(2-methoxyethoxy)methyl, benzyloxymethyl,beta-(trimethylsilyl)ethoxymethyl, and tetrahydropyranyl ethers.Examples of arylalkyl ethers include benzyl, p-methoxybenzyl (MPM),3,4-dimethoxybenzyl, O-nitrobenzyl, p-nitrobenzyl, p-halobenzyl,2,6-dichlorobenzyl, p-cyanobenzyl, and 2- and 4-picolyl.

Amino protecting groups are well known in the art and include thosedescribed in detail in Protecting Groups in Organic Synthesis, T. W.Greene and P. G. M. Wuts, 3^(rd) edition, John Wiley & Sons, 1999, andPhilip Kocienski, in “Protecting Groups”, Georg Thieme Verlag Stuttgart,New York, 1994, the entireties of which is incorporated herein byreference. Suitable amino protecting groups include, but are not limitedto, aralkylamines, carbamates, cyclic imides, allyl amines, amides, andthe like. Examples of such groups include t-butyloxycarbonyl (BOC),ethyloxycarbonyl, methyloxycarbonyl, trichloroethyloxycarbonyl,allyloxycarbonyl (Alloc), benzyloxocarbonyl (CBZ), allyl, phthalimide,benzyl (Bn), fluorenylmethylcarbonyl (Fmoc), formyl, acetyl,chloroacetyl, dichloroacetyl, trichloroacetyl, phenylacetyl,trifluoroacetyl, benzoyl, and the like.

One of skill in the art will appreciate that various functional groupspresent in compounds of the invention such as aliphatic groups,alcohols, carboxylic acids, esters, amides, aldehydes, halogens andnitriles can be interconverted by techniques well known in the artincluding, but not limited to reduction, oxidation, esterification,hydrolysis, partial oxidation, partial reduction, halogenation,dehydration, partial hydration, and hydration. See, for example,“March's Advanced Organic Chemistry”, 5^(th) Ed., Ed.: Smith, M. B. andMarch, J., John Wiley & Sons, New York: 2001, the entirety of which isincorporated herein by reference. Such interconversions may require oneor more of the aforementioned techniques, and certain methods forsynthesizing compounds of the invention are described below.

In one aspect, certain compounds of the present invention of Formula I,or subformulae thereof, are generally prepared according to Scheme 1 setforth below:

In Scheme 1 above, PG is a nitrogen protecting group, and each of R¹,R², R³, R⁴, R⁵, Ring A, m, n, and p is as defined above and described inembodiments herein, both singly and in combination.

As shown generally in Scheme 1, an aldehyde according to structure A maybe condensed with a ketone such as acetone in the presence of a base toyield intermediate B, for example by following General Procedures E orF. The General Procedures are described in more detail in theExemplification, below. Condensation with an amine such as NH₂R³, e.g.methylamine, and an aldehyde of structure C, provides compounds ofstructure D. In some embodiments, such compounds are CXCR4 inhibitorsaccording to the present invention. In other embodiments, compounds ofstructure D are reduced according to General Procedure A to providecompounds of structure E. In compounds of structure F where R² is anappropriate leaving group (LG), cross-coupling (such as Pd-catalyzedcoupling) may be performed to provide compounds of structure G. If R² ishydrogen in structure F, halogenation or formation of a leaving groupsuch as triflate precedes the coupling reaction. Alternatively, if R² ishydrogen in structure F, alkylation such as a formylation withparaformaldehyde or DMF may be used to provide certain compounds ofstructure G.

Alternatively, as shown in Scheme 2, piperidone compounds of structure Hmay be reduced according to General Procedure A to afford compounds ofstructure I and subsequently reacted with an appropriate electrophile offormula LG-R³, wherein LG refers to an appropriate leaving group such ashalide or mesylate, affording compounds of structure J.

5. Uses, Formulation and Administration, and Co-Administered AdditionalTherapeutic Agents Pharmaceutically Acceptable Compositions

According to another embodiment, the invention provides a compositioncomprising a compound of this invention or a pharmaceutically acceptablederivative thereof and a pharmaceutically acceptable carrier, adjuvant,or vehicle. The amount of compound in compositions of this invention issuch that is effective to measurably inhibit CXCR4, or a mutant thereof,in a biological sample or in a patient. In certain embodiments, theamount of compound in compositions of this invention is such that iseffective to measurably inhibit CXCR4, or a mutant thereof, in abiological sample or in a patient. In certain embodiments, a compositionof this invention is formulated for administration to a patient in needof such composition. In some embodiments, a composition of thisinvention is formulated for oral administration to a patient.

The term “patient,” as used herein, means an animal, preferably amammal, and most preferably a human.

The term “pharmaceutically acceptable carrier, adjuvant, or vehicle”refers to a non-toxic carrier, adjuvant, or vehicle that does notdestroy the pharmacological activity of the compound with which it isformulated. Pharmaceutically acceptable carriers, adjuvants or vehiclesthat may be used in the compositions of this invention include, but arenot limited to, ion exchangers, alumina, aluminum stearate, lecithin,serum proteins, such as human serum albumin, buffer substances such asphosphates, glycine, sorbic acid, potassium sorbate, partial glyceridemixtures of saturated vegetable fatty acids, water, salts orelectrolytes, such as protamine sulfate, disodium hydrogen phosphate,potassium hydrogen phosphate, sodium chloride, zinc salts, colloidalsilica, magnesium trisilicate, polyvinyl pyrrolidone, cellulose-basedsubstances, polyethylene glycol, sodium carboxymethylcellulose,polyacrylates, waxes, polyethylene-polyoxypropylene-block polymers,polyethylene glycol and wool fat.

A “pharmaceutically acceptable derivative” means any non-toxic salt,ester, salt of an ester or other derivative of a compound of thisinvention that, upon administration to a recipient, is capable ofproviding, either directly or indirectly, a compound of this inventionor an inhibitorily active metabolite or residue thereof.

As used herein, the term “inhibitorily active metabolite or residuethereof” means that a metabolite or residue thereof is also an inhibitorof CXCR4, or a mutant thereof.

Compositions of the present invention may be administered orally,parenterally, by inhalation spray, topically, rectally, nasally,buccally, vaginally or via an implanted reservoir. The term “parenteral”as used herein includes subcutaneous, intravenous, intramuscular,intra-articular, intra-synovial, intrasternal, intrathecal,intrahepatic, intralesional and intracranial injection or infusiontechniques. Preferably, the compositions are administered orally,intraperitoneally or intravenously. Sterile injectable forms of thecompositions of this invention may be aqueous or oleaginous suspension.These suspensions may be formulated according to techniques known in theart using suitable dispersing or wetting agents and suspending agents.The sterile injectable preparation may also be a sterile injectablesolution or suspension in a non-toxic parenterally acceptable diluent orsolvent, for example as a solution in 1,3-butanediol. Among theacceptable vehicles and solvents that may be employed are water,Ringer's solution and isotonic sodium chloride solution. In addition,sterile, fixed oils are conventionally employed as a solvent orsuspending medium.

For this purpose, any bland fixed oil may be employed includingsynthetic mono- or di-glycerides. Fatty acids, such as oleic acid andits glyceride derivatives are useful in the preparation of injectables,as are natural pharmaceutically-acceptable oils, such as olive oil orcastor oil, especially in their polyoxyethylated versions. These oilsolutions or suspensions may also contain a long-chain alcohol diluentor dispersant, such as carboxymethyl cellulose or similar dispersingagents that are commonly used in the formulation of pharmaceuticallyacceptable dosage forms including emulsions and suspensions. Othercommonly used surfactants, such as Tweens, Spans and other emulsifyingagents or bioavailability enhancers which are commonly used in themanufacture of pharmaceutically acceptable solid, liquid, or otherdosage forms may also be used for the purposes of formulation.

Pharmaceutically acceptable compositions of this invention may be orallyadministered in any orally acceptable dosage form including, but notlimited to, capsules, tablets, aqueous suspensions or solutions. In thecase of tablets for oral use, carriers commonly used include lactose andcorn starch. Lubricating agents, such as magnesium stearate, are alsotypically added. For oral administration in a capsule form, usefuldiluents include lactose and dried cornstarch. When aqueous suspensionsare required for oral use, the active ingredient is combined withemulsifying and suspending agents. If desired, certain sweetening,flavoring or coloring agents may also be added.

Alternatively, pharmaceutically acceptable compositions of thisinvention may be administered in the form of suppositories for rectaladministration. These can be prepared by mixing the agent with asuitable non-irritating excipient that is solid at room temperature butliquid at rectal temperature and therefore will melt in the rectum torelease the drug. Such materials include cocoa butter, beeswax andpolyethylene glycols.

Pharmaceutically acceptable compositions of this invention may also beadministered topically, especially when the target of treatment includesareas or organs readily accessible by topical application, includingdiseases of the eye, the skin, or the lower intestinal tract. Suitabletopical formulations are readily prepared for each of these areas ororgans.

Topical application for the lower intestinal tract can be effected in arectal suppository formulation (see above) or in a suitable enemaformulation. Topically-transdermal patches may also be used.

For topical applications, provided pharmaceutically acceptablecompositions may be formulated in a suitable ointment containing theactive component suspended or dissolved in one or more carriers.Carriers for topical administration of compounds of this inventioninclude, but are not limited to, mineral oil, liquid petrolatum, whitepetrolatum, propylene glycol, polyoxyethylene, polyoxypropylenecompound, emulsifying wax and water. Alternatively, providedpharmaceutically acceptable compositions can be formulated in a suitablelotion or cream containing the active components suspended or dissolvedin one or more pharmaceutically acceptable carriers. Suitable carriersinclude, but are not limited to, mineral oil, sorbitan monostearate,polysorbate 60, cetyl esters wax, cetearyl alcohol, 2-octyldodecanol,benzyl alcohol and water.

For ophthalmic use, provided pharmaceutically acceptable compositionsmay be formulated as micronized suspensions in isotonic, pH adjustedsterile saline, or, preferably, as solutions in isotonic, pH adjustedsterile saline, either with or without a preservative such asbenzylalkonium chloride. Alternatively, for ophthalmic uses, thepharmaceutically acceptable compositions may be formulated in anointment such as petrolatum.

Pharmaceutically acceptable compositions of this invention may also beadministered by nasal aerosol or inhalation. Such compositions areprepared according to techniques well-known in the art of pharmaceuticalformulation and may be prepared as solutions in saline, employing benzylalcohol or other suitable preservatives, absorption promoters to enhancebioavailability, fluorocarbons, and/or other conventional solubilizingor dispersing agents.

Most preferably, pharmaceutically acceptable compositions of thisinvention are formulated for oral administration. Such formulations maybe administered with or without food. In some embodiments,pharmaceutically acceptable compositions of this invention areadministered without food. In other embodiments, pharmaceuticallyacceptable compositions of this invention are administered with food.

The amount of compounds of the present invention that may be combinedwith the carrier materials to produce a composition in a single dosageform will vary depending upon the host treated, the particular mode ofadministration. Preferably, provided compositions should be formulatedso that a dosage of between 0.01-100 mg/kg body weight/day of theinhibitor can be administered to a patient receiving these compositions.

It should also be understood that a specific dosage and treatmentregimen for any particular patient will depend upon a variety offactors, including the activity of the specific compound employed, theage, body weight, general health, sex, diet, time of administration,rate of excretion, drug combination, and the judgment of the treatingphysician and the severity of the particular disease being treated. Theamount of a compound of the present invention in the composition willalso depend upon the particular compound in the composition.

Uses of Compounds and Pharmaceutically Acceptable Compositions

Compounds and compositions described herein are generally useful for theinhibition of CXCR4 or a mutant thereof.

The activity of a compound utilized in this invention as an inhibitor ofCXCR4, or a mutant thereof, may be assayed in vitro, in vivo or in acell line. In vitro assays include assays that determine inhibition ofCXCR4, or a mutant thereof. Alternate in vitro assays quantitate theability of the inhibitor to bind to CXCR4. Detailed conditions forassaying a compound utilized in this invention as an inhibitor of CXCR4,or a mutant thereof, are set forth in the Examples below.

As used herein, the terms “treatment,” “treat,” and “treating” refer toreversing, alleviating, delaying the onset of, or inhibiting theprogress of a disease or disorder, or one or more symptoms thereof, asdescribed herein. In some embodiments, treatment may be administeredafter one or more symptoms have developed. In other embodiments,treatment may be administered in the absence of symptoms. For example,treatment may be administered to a susceptible individual prior to theonset of symptoms (e.g., in light of a history of symptoms and/or inlight of genetic or other susceptibility factors). Treatment may also becontinued after symptoms have resolved, for example to prevent or delaytheir recurrence.

Provided compounds are inhibitors of CXCR4 and are therefore useful fortreating one or more disorders associated with activity of CXCR4. Thus,in certain embodiments, the present invention provides a method fortreating a CXCR4-mediated disorder comprising the step of administeringto a patient in need thereof a compound of the present invention, orpharmaceutically acceptable composition thereof.

As used herein, the terms “CXCR4-mediated” disorders, diseases, and/orconditions as used herein means any disease or other deleteriouscondition in which CXCR4, or a mutant thereof, is known to play a role.Accordingly, another embodiment of the present invention relates totreating or lessening the severity of one or more diseases in whichCXCR4, or a mutant thereof, are known to play a role.

In some embodiments, the present invention provides a method fortreating one or more disorders, diseases, and/or conditions wherein thedisorder, disease, or condition includes, but is not limited to, acellular proliferative disorder.

Cellular Proliferative Disorders

The present invention features methods and compositions for thediagnosis and prognosis of cellular proliferative disorders (e.g.,cancer) and the treatment of these disorders by targeting CXCR4.Cellular proliferative disorders described herein include, e.g., cancer,obesity, and proliferation-dependent diseases. Such disorders may bediagnosed using methods known in the art.

Cancer

Cancer includes, in one embodiment, without limitation, leukemias (e.g.,acute leukemia, acute lymphocytic leukemia, acute myelocytic leukemia,acute myeloblastic leukemia, acute promyelocytic leukemia, acutemyelomonocytic leukemia, acute monocytic leukemia, acuteerythroleukemia, chronic leukemia, chronic myelocytic leukemia, chroniclymphocytic leukemia), polycythemia vera, lymphoma (e.g., Hodgkin'sdisease or non-Hodgkin's disease), Waldenstrom's macroglobulinemia,multiple myeloma, heavy chain disease, and solid tumors such as sarcomasand carcinomas (e.g., fibrosarcoma, myxosarcoma, liposarcoma,chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma,endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma,synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma,rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer,ovarian cancer, prostate cancer, squamous cell carcinoma, basal cellcarcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous glandcarcinoma, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterinecancer, testicular cancer, lung carcinoma, small cell lung carcinoma,bladder carcinoma, epithelial carcinoma, glioma, astrocytoma,glioblastoma multiforme (GBM, also known as glioblastoma),medulloblastoma, craniopharyngioma, ependymoma, pinealoma,hemangioblastoma, acoustic neuroma, oligodendroglioma, schwannoma,neurofibrosarcoma, meningioma, melanoma, neuroblastoma, andretinoblastoma).

In some embodiments, the cancer is glioma, astrocytoma, glioblastomamultiforme (GBM, also known as glioblastoma), medulloblastoma,craniopharyngioma, ependymoma, pinealoma, hemangioblastoma, acousticneuroma, oligodendroglioma, schwannoma, neurofibrosarcoma, meningioma,melanoma, neuroblastoma, or retinoblastoma.

In some embodiments, the cancer is acoustic neuroma, astrocytoma (e.g.Grade I—Pilocytic Astrocytoma, Grade II—Low-grade Astrocytoma, GradeIII—Anaplastic Astrocytoma, or Grade IV—Glioblastoma (GBM)), chordoma,CNS lymphoma, craniopharyngioma, brain stem glioma, ependymoma, mixedglioma, optic nerve glioma, subependymoma, medulloblastoma, meningioma,metastatic brain tumor, oligodendroglioma, pituitary tumors, primitiveneuroectodermal (PNET) tumor, or schwannoma. In some embodiments, thecancer is a type found more commonly in children than adults, such asbrain stem glioma, craniopharyngioma, ependymoma, juvenile pilocyticastrocytoma (JPA), medulloblastoma, optic nerve glioma, pineal tumor,primitive neuroectodermal tumors (PNET), or rhabdoid tumor. In someembodiments, the patient is an adult human. In some embodiments, thepatient is a child or pediatric patient.

Cancer includes, in another embodiment, without limitation,mesothelioma, hepatobilliary (hepatic and billiary duct), bone cancer,pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous orintraocular melanoma, ovarian cancer, colon cancer, rectal cancer,cancer of the anal region, stomach cancer, gastrointestinal (gastric,colorectal, and duodenal), uterine cancer, carcinoma of the fallopiantubes, carcinoma of the endometrium, carcinoma of the cervix, carcinomaof the vagina, carcinoma of the vulva, Hodgkin's Disease, cancer of theesophagus, cancer of the small intestine, cancer of the endocrinesystem, cancer of the thyroid gland, cancer of the parathyroid gland,cancer of the adrenal gland, sarcoma of soft tissue, cancer of theurethra, cancer of the penis, prostate cancer, testicular cancer,chronic or acute leukemia, chronic myeloid leukemia, lymphocyticlymphomas, cancer of the bladder, cancer of the kidney or ureter, renalcell carcinoma, carcinoma of the renal pelvis, non-Hodgkins's lymphoma,spinal axis tumors, brain stem glioma, pituitary adenoma, adrenocorticalcancer, gall bladder cancer, multiple myeloma, cholangiocarcinoma,fibrosarcoma, neuroblastoma, retinoblastoma, or a combination of one ormore of the foregoing cancers.

In some embodiments, the cancer is selected from hepatocellularcarcinoma, ovarian cancer, ovarian epithelial cancer, or fallopian tubecancer; papillary serous cystadenocarcinoma or uterine papillary serouscarcinoma (UPSC); prostate cancer; testicular cancer; gallbladdercancer; hepatocholangiocarcinoma; soft tissue and bone synovial sarcoma;rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewing sarcoma;anaplastic thyroid cancer; adrenocortical adenoma; pancreatic cancer;pancreatic ductal carcinoma or pancreatic adenocarcinoma;gastrointestinal/stomach (GIST) cancer; lymphoma; squamous cellcarcinoma of the head and neck (SCCHN); salivary gland cancer; glioma,or brain cancer; neurofibromatosis-1 associated malignant peripheralnerve sheath tumors (MPNST); Waldenstrom's macroglobulinemia; ormedulloblastoma.

In some embodiments, the cancer is selected from hepatocellularcarcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovariancancer, ovarian epithelial cancer, fallopian tube cancer, papillaryserous cystadenocarcinoma, uterine papillary serous carcinoma (UPSC),hepatocholangiocarcinoma, soft tissue and bone synovial sarcoma,rhabdomyosarcoma, osteosarcoma, anaplastic thyroid cancer,adrenocortical adenoma, pancreatic cancer, pancreatic ductal carcinoma,pancreatic adenocarcinoma, glioma, neurofibromatosis-1 associatedmalignant peripheral nerve sheath tumors (MPNST), Waldenstrom'smacroglobulinemia, or medulloblastoma.

In some embodiments, the present invention provides a method fortreating a cancer that presents as a solid tumor, such as a sarcoma,carcinoma, or lymphoma, comprising the step of administering a disclosedcompound, or a pharmaceutically acceptable salt thereof, to a patient inneed thereof. Solid tumors generally comprise an abnormal mass of tissuethat typically does not include cysts or liquid areas. In someembodiments, the cancer is selected from renal cell carcinoma, or kidneycancer; hepatocellular carcinoma (HCC) or hepatoblastoma, or livercancer; melanoma; breast cancer; colorectal carcinoma, or colorectalcancer; colon cancer; rectal cancer; anal cancer; lung cancer, such asnon-small cell lung cancer (NSCLC) or small cell lung cancer (SCLC);ovarian cancer, ovarian epithelial cancer, ovarian carcinoma, orfallopian tube cancer; papillary serous cystadenocarcinoma or uterinepapillary serous carcinoma (UPSC); prostate cancer; testicular cancer;gallbladder cancer; hepatocholangiocarcinoma; soft tissue and bonesynovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma; Ewingsarcoma; anaplastic thyroid cancer; adrenocortical carcinoma; pancreaticcancer; pancreatic ductal carcinoma or pancreatic adenocarcinoma;gastrointestinal/stomach (GIST) cancer; lymphoma; squamous cellcarcinoma of the head and neck (SCCHN); salivary gland cancer; glioma,or brain cancer; neurofibromatosis-1 associated malignant peripheralnerve sheath tumors (MPNST); Waldenstrom's macroglobulinemia; ormedulloblastoma.

In some embodiments, the cancer is selected from renal cell carcinoma,hepatocellular carcinoma (HCC), hepatoblastoma, colorectal carcinoma,colorectal cancer, colon cancer, rectal cancer, anal cancer, ovariancancer, ovarian epithelial cancer, ovarian carcinoma, fallopian tubecancer, papillary serous cystadenocarcinoma, uterine papillary serouscarcinoma (UPSC), hepatocholangiocarcinoma, soft tissue and bonesynovial sarcoma, rhabdomyosarcoma, osteosarcoma, chondrosarcoma,anaplastic thyroid cancer, adrenocortical carcinoma, pancreatic cancer,pancreatic ductal carcinoma, pancreatic adenocarcinoma, glioma, braincancer, neurofibromatosis-1 associated malignant peripheral nerve sheathtumors (MPNST), Waldenstrom's macroglobulinemia, or medulloblastoma.

In some embodiments, the cancer is selected from hepatocellularcarcinoma (HCC), hepatoblastoma, colon cancer, rectal cancer, ovariancancer, ovarian epithelial cancer, ovarian carcinoma, fallopian tubecancer, papillary serous cystadenocarcinoma, uterine papillary serouscarcinoma (UPSC), hepatocholangiocarcinoma, soft tissue and bonesynovial sarcoma, rhabdomyosarcoma, osteosarcoma, anaplastic thyroidcancer, adrenocortical carcinoma, pancreatic cancer, pancreatic ductalcarcinoma, pancreatic adenocarcinoma, glioma, neurofibromatosis-1associated malignant peripheral nerve sheath tumors (MPNST),Waldenstrom's macroglobulinemia, or medulloblastoma.

In some embodiments, the cancer is hepatocellular carcinoma (HCC). Insome embodiments, the cancer is hepatoblastoma. In some embodiments, thecancer is colon cancer. In some embodiments, the cancer is rectalcancer. In some embodiments, the cancer is ovarian cancer, or ovariancarcinoma. In some embodiments, the cancer is ovarian epithelial cancer.In some embodiments, the cancer is fallopian tube cancer. In someembodiments, the cancer is papillary serous cystadenocarcinoma. In someembodiments, the cancer is uterine papillary serous carcinoma (UPSC). Insome embodiments, the cancer is hepatocholangiocarcinoma. In someembodiments, the cancer is soft tissue and bone synovial sarcoma. Insome embodiments, the cancer is rhabdomyosarcoma. In some embodiments,the cancer is osteosarcoma. In some embodiments, the cancer isanaplastic thyroid cancer. In some embodiments, the cancer isadrenocortical carcinoma. In some embodiments, the cancer is pancreaticcancer, or pancreatic ductal carcinoma. In some embodiments, the canceris pancreatic adenocarcinoma. In some embodiments, the cancer is glioma.In some embodiments, the cancer is malignant peripheral nerve sheathtumors (MPNST). In some embodiments, the cancer is neurofibromatosis-1associated MPNST. In some embodiments, the cancer is Waldenstrom'smacroglobulinemia. In some embodiments, the cancer is medulloblastoma.

The present invention further features methods and compositions for thediagnosis, prognosis and treatment of viral-associated cancers,including human immunodeficiency virus (HIV) associated solid tumors,human papilloma virus (HPV)-16 positive incurable solid tumors, andadult T-cell leukemia, which is caused by human T-cell leukemia virustype I (HTLV-I) and is a highly aggressive form of CD4+ T-cell leukemiacharacterized by clonal integration of HTLV-I in leukemic cells (Seehttps://clinicaltrials.gov/ct2/show/study/NCT02631746); as well asvirus-associated tumors in gastric cancer, nasopharyngeal carcinoma,cervical cancer, vaginal cancer, vulvar cancer, squamous cell carcinomaof the head and neck, and Merkel cell carcinoma. (Seehttps://clinicaltrials.gov/ct2/show/study/NCT02488759; see alsohttps://clinicaltrials.gov/ct2/show/study/NC TO240886;https://clinicaltrials.gov/ct2/show/NCT02426892)

In some embodiments, the present invention provides a method fortreating a tumor in a patient in need thereof, comprising administeringto the patient any of the compounds, salts or pharmaceuticalcompositions described herein. In some embodiments, the tumor comprisesany of the cancers described herein. In some embodiments, the tumorcomprises melanoma cancer. In some embodiments, the tumor comprisesbreast cancer. In some embodiments, the tumor comprises lung cancer. Insome embodiments the tumor comprises small cell lung cancer (SCLC). Insome embodiments the tumor comprises non-small cell lung cancer (NSCLC).

In some embodiments, the tumor is treated by arresting further growth ofthe tumor. In some embodiments, the tumor is treated by reducing thesize (e.g., volume or mass) of the tumor by at least 5%, 10%, 25%, 50%,75%, 90% or 99% relative to the size of the tumor prior to treatment. Insome embodiments, tumors are treated by reducing the quantity of thetumors in the patient by at least 5%, 10%, 25%, 50%, 75%, 90% or 99%relative to the quantity of tumors prior to treatment.

Primary Immune Deficiencies

In some embodiments, the present invention provides a method fortreating one or more disorders, diseases, and/or conditions wherein thedisorder, disease, or condition includes, but is not limited to, aprimary immunodeficiency disease or disorder, comprising administeringto a patient in need thereof an effective amount of a disclosedcompound. Primary immune deficiencies treatable by the methods of thepresent invention include: warts, hypogammaglobulinemia, infections,myelokathexis (WHIMs) syndrome; severe congenital neutropenia (SCN),especially those arising from G6PC3 deficiency (McDermott et al. (2010)Blood 116:2793-2802); GATA2 deficiency (Mono MAC syndrome)(Maciejweski-Duval et al. (2015) J. Leukoc. Biol. 5MA0815-288R (Epub.ahead of printing); idiopathic CD4+ T lymphocytopenia (ICL); andWiskott-Aldrich Syndrome.

The compounds and compositions, according to the method of the presentinvention, may be administered using any amount and any route ofadministration effective for treating or lessening the severity of acancer, an autoimmune disorder, a primary immune deficiency, aproliferative disorder, an inflammatory disorder, a neurodegenerative orneurological disorder, schizophrenia, a bone-related disorder, liverdisease, or a cardiac disorder. The exact amount required will vary fromsubject to subject, depending on the species, age, and general conditionof the subject, the severity of the disease or disorder, the particularagent, its mode of administration, and the like. Compounds of theinvention are preferably formulated in dosage unit form for ease ofadministration and uniformity of dosage. The expression “dosage unitform” as used herein refers to a physically discrete unit of agentappropriate for the patient to be treated. It will be understood,however, that the total daily usage of the compounds and compositions ofthe present invention will be decided by the attending physician withinthe scope of sound medical judgment. The specific effective dose levelfor any particular patient or organism will depend upon a variety offactors including the disorder being treated and the severity of thedisorder; the activity of the specific compound employed; the specificcomposition employed; the age, body weight, general health, sex and dietof the patient; the time of administration, route of administration, andrate of excretion of the specific compound employed; the duration of thetreatment; drugs used in combination or coincidental with the specificcompound employed, and like factors well known in the medical arts. Theterm “patient”, as used herein, means an animal, preferably a mammal,and most preferably a human.

Pharmaceutically acceptable compositions of this invention can beadministered to humans and other animals orally, rectally, parenterally,intracisternally, intravaginally, intraperitoneally, topically (as bypowders, ointments, or drops), bucally, as an oral or nasal spray, orthe like, depending on the severity of the disease or disorder beingtreated. In certain embodiments, the compounds of the invention may beadministered orally or parenterally at dosage levels of about 0.01 mg/kgto about 50 mg/kg and preferably from about 1 mg/kg to about 25 mg/kg,of subject body weight per day, one or more times a day, to obtain thedesired therapeutic effect.

Liquid dosage forms for oral administration include, but are not limitedto, pharmaceutically acceptable emulsions, microemulsions, solutions,suspensions, syrups and elixirs. In addition to the active compounds,the liquid dosage forms may contain inert diluents commonly used in theart such as, for example, water or other solvents, solubilizing agentsand emulsifiers such as ethyl alcohol, isopropyl alcohol, ethylcarbonate, ethyl acetate, benzyl alcohol, benzyl benzoate, propyleneglycol, 1,3-butylene glycol, dimethylformamide, oils (in particular,cottonseed, groundnut, corn, germ, olive, castor, and sesame oils),glycerol, tetrahydrofurfuryl alcohol, polyethylene glycols and fattyacid esters of sorbitan, and mixtures thereof. Besides inert diluents,the oral compositions can also include adjuvants such as wetting agents,emulsifying and suspending agents, sweetening, flavoring, and perfumingagents.

Injectable preparations, for example, sterile injectable aqueous oroleaginous suspensions may be formulated according to the known artusing suitable dispersing or wetting agents and suspending agents. Thesterile injectable preparation may also be a sterile injectablesolution, suspension or emulsion in a nontoxic parenterally acceptablediluent or solvent, for example, as a solution in 1,3-butanediol. Amongthe acceptable vehicles and solvents that may be employed are water,Ringer's solution, U.S.P. and isotonic sodium chloride solution. Inaddition, sterile, fixed oils are conventionally employed as a solventor suspending medium. For this purpose any bland fixed oil can beemployed including synthetic mono- or diglycerides. In addition, fattyacids such as oleic acid are used in the preparation of injectables.

Injectable formulations can be sterilized, for example, by filtrationthrough a bacterial-retaining filter, or by incorporating sterilizingagents in the form of sterile solid compositions which can be dissolvedor dispersed in sterile water or other sterile injectable medium priorto use.

In order to prolong the effect of a compound of the present invention,it is often desirable to slow the absorption of the compound fromsubcutaneous or intramuscular injection. This may be accomplished by theuse of a liquid suspension of crystalline or amorphous material withpoor water solubility. The rate of absorption of the compound thendepends upon its rate of dissolution that, in turn, may depend uponcrystal size and crystalline form. Alternatively, delayed absorption ofa parenterally administered compound form is accomplished by dissolvingor suspending the compound in an oil vehicle. Injectable depot forms aremade by forming microencapsule matrices of the compound in biodegradablepolymers such as polylactide-polyglycolide. Depending upon the ratio ofcompound to polymer and the nature of the particular polymer employed,the rate of compound release can be controlled. Examples of otherbiodegradable polymers include poly(orthoesters) and poly(anhydrides).Depot injectable formulations are also prepared by entrapping thecompound in liposomes or microemulsions that are compatible with bodytissues.

Compositions for rectal or vaginal administration are preferablysuppositories which can be prepared by mixing the compounds of thisinvention with suitable non-irritating excipients or carriers such ascocoa butter, polyethylene glycol or a suppository wax which are solidat ambient temperature but liquid at body temperature and therefore meltin the rectum or vaginal cavity and release the active compound.

Solid dosage forms for oral administration include capsules, tablets,pills, powders, and granules. In such solid dosage forms, the activecompound is mixed with at least one inert, pharmaceutically acceptableexcipient or carrier such as sodium citrate or dicalcium phosphateand/or a) fillers or extenders such as starches, lactose, sucrose,glucose, mannitol, and silicic acid, b) binders such as, for example,carboxymethylcellulose, alginates, gelatin, polyvinylpyrrolidinone,sucrose, and acacia, c) humectants such as glycerol, d) disintegratingagents such as agar-agar, calcium carbonate, potato or tapioca starch,alginic acid, certain silicates, and sodium carbonate, e) solutionretarding agents such as paraffin, f) absorption accelerators such asquaternary ammonium compounds, g) wetting agents such as, for example,cetyl alcohol and glycerol monostearate, h) absorbents such as kaolinand bentonite clay, and i) lubricants such as talc, calcium stearate,magnesium stearate, solid polyethylene glycols, sodium lauryl sulfate,and mixtures thereof. In the case of capsules, tablets and pills, thedosage form may also comprise buffering agents.

Solid compositions of a similar type may also be employed as fillers insoft and hard-filled gelatin capsules using such excipients as lactoseor milk sugar as well as high molecular weight polyethylene glycols andthe like. The solid dosage forms of tablets, dragees, capsules, pills,and granules can be prepared with coatings and shells such as entericcoatings and other coatings well known in the pharmaceutical formulatingart. They may optionally contain opacifying agents and can also be of acomposition that they release the active ingredient(s) only, orpreferentially, in a certain part of the intestinal tract, optionally,in a delayed manner. Examples of embedding compositions that can be usedinclude polymeric substances and waxes. Solid compositions of a similartype may also be employed as fillers in soft and hard-filled gelatincapsules using such excipients as lactose or milk sugar as well as highmolecular weight polethylene glycols and the like.

The active compounds can also be in micro-encapsulated form with one ormore excipients as noted above. The solid dosage forms of tablets,dragees, capsules, pills, and granules can be prepared with coatings andshells such as enteric coatings, release controlling coatings and othercoatings well known in the pharmaceutical formulating art. In such soliddosage forms the active compound may be admixed with at least one inertdiluent such as sucrose, lactose or starch. Such dosage forms may alsocomprise, as is normal practice, additional substances other than inertdiluents, e.g., tableting lubricants and other tableting aids such amagnesium stearate and microcrystalline cellulose. In the case ofcapsules, tablets and pills, the dosage forms may also comprisebuffering agents. They may optionally contain opacifying agents and canalso be of a composition that they release the active ingredient(s)only, or preferentially, in a certain part of the intestinal tract,optionally, in a delayed manner. Examples of embedding compositions thatcan be used include polymeric substances and waxes.

Dosage forms for topical or transdermal administration of a compound ofthis invention include ointments, pastes, creams, lotions, gels,powders, solutions, sprays, inhalants or patches. The active componentis admixed under sterile conditions with a pharmaceutically acceptablecarrier and any needed preservatives or buffers as may be required.Ophthalmic formulation, ear drops, and eye drops are also contemplatedas being within the scope of this invention. Additionally, the presentinvention contemplates the use of transdermal patches, which have theadded advantage of providing controlled delivery of a compound to thebody. Such dosage forms can be made by dissolving or dispensing thecompound in the proper medium. Absorption enhancers can also be used toincrease the flux of the compound across the skin. The rate can becontrolled by either providing a rate controlling membrane or bydispersing the compound in a polymer matrix or gel.

According to one embodiment, the invention relates to a method ofinhibiting CXCR4 activity in a biological sample comprising the step ofcontacting said biological sample with a compound of this invention, ora composition comprising said compound.

According to another embodiment, the invention relates to a method ofinhibiting CXCR4, or a mutant thereof, activity in a biological samplecomprising the step of contacting said biological sample with a compoundof this invention, or a composition comprising said compound. In certainembodiments, the invention relates to a method of irreversiblyinhibiting CXCR4, or a mutant thereof, activity in a biological samplecomprising the step of contacting said biological sample with a compoundof this invention, or a composition comprising said compound.

The term “biological sample”, as used herein, includes, withoutlimitation, cell cultures or extracts thereof; biopsied materialobtained from a mammal or extracts thereof; and blood, saliva, urine,feces, semen, tears, or other body fluids or extracts thereof.

Another embodiment of the present invention relates to a method ofinhibiting CXCR4 in a patient comprising the step of administering tosaid patient a compound of the present invention, or a compositioncomprising said compound.

According to another embodiment, the invention relates to a method ofinhibiting CXCR4, or a mutant thereof, activity in a patient comprisingthe step of administering to said patient a compound of the presentinvention, or a composition comprising said compound. According tocertain embodiments, the invention relates to a method of irreversiblyinhibiting CXCR4, or a mutant thereof, activity in a patient comprisingthe step of administering to said patient a compound of the presentinvention, or a composition comprising said compound. In otherembodiments, the present invention provides a method for treating adisorder mediated by CXCR4, or a mutant thereof, in a patient in needthereof, comprising the step of administering to said patient a compoundaccording to the present invention or pharmaceutically acceptablecomposition thereof. Such disorders are described in detail herein.

Co Administration of Additional Therapeutic Agents

Depending upon the particular condition, or disease, to be treated,additional therapeutic agents that are normally administered to treatthat condition, may also be present in the compositions of thisinvention. As used herein, additional therapeutic agents that arenormally administered to treat a particular disease, or condition, areknown as “appropriate for the disease, or condition, being treated.”

In some embodiments, the present invention provides a method of treatinga disclosed disease or condition comprising administering to a patientin need thereof an effective amount of a compound disclosed herein or apharmaceutically acceptable salt thereof and co-administeringsimultaneously or sequentially an effective amount of one or moreadditional therapeutic agents, such as those described herein. In someembodiments, the method includes co-administering one additionaltherapeutic agent. In some embodiments, the method includesco-administering two additional therapeutic agents. In some embodiments,the combination of the disclosed compound and the additional therapeuticagent or agents acts synergistically.

In some embodiments, the additional therapeutic agent is selected froman immunostimulatory therapeutic compound. In some embodiments, theimmunostimulatory therapeutic compound is selected from elotuzumab,mifamurtide, an agonist or activator of a toll-like receptor, or anactivator of RORyt.

In some embodiments, the method further comprises administering to saidpatient a third therapeutic agent, such as an immune checkpointinhibitor. In some embodiments, the method comprises administering tothe patient in need thereof three therapeutic agents selected from acompound disclosed herein or a pharmaceutically acceptable salt thereof,an immunostimulatory therapeutic compound, and an immune checkpointinhibitor.

Other checkpoint inhibitors that may be used in the present inventioninclude OX40 agonists. OX40 agonists that are being studied in clinicaltrials include PF-04518600/PF-8600 (Pfizer), an agonistic anti-OX40antibody, in metastatic kidney cancer (NCT03092856) and advanced cancersand neoplasms (NCT02554812; NCT05082566); GSK3174998 (Merck), anagonistic anti-OX40 antibody, in Phase 1 cancer trials (NCT02528357);MEDI0562 (Medimmune/AstraZeneca), an agonistic anti-OX40 antibody, inadvanced solid tumors (NCT02318394 and NCT02705482); MEDI6469, anagonistic anti-OX40 antibody (Medimmune/AstraZeneca), in patients withcolorectal cancer (NCT02559024), breast cancer (NCT01862900), head andneck cancer (NCT02274155) and metastatic prostate cancer (NCT01303705);and BMS-986178 (Bristol-Myers Squibb) an agonistic anti-OX40 antibody,in advanced cancers (NCT02737475).

Other checkpoint inhibitors that may be used in the present inventioninclude CD137 (also called 4-1BB) agonists. CD137 agonists that arebeing studied in clinical trials include utomilumab (PF-05082566,Pfizer) an agonistic anti-CD137 antibody, in diffuse large B-celllymphoma (NCT02951156) and in advanced cancers and neoplasms(NCT02554812 and NCT05082566); urelumab (BMS-663513, Bristol-MyersSquibb), an agonistic anti-CD137 antibody, in melanoma and skin cancer(NCT02652455) and glioblastoma and gliosarcoma (NC T02658981).

Other checkpoint inhibitors that may be used in the present inventioninclude CD27 agonists. CD27 agonists that are being studied in clinicaltrials include varlilumab (CDX-1127, Celldex Therapeutics) an agonisticanti-CD27 antibody, in squamous cell head and neck cancer, ovariancarcinoma, colorectal cancer, renal cell cancer, and glioblastoma(NCT02335918); lymphomas (NCT01460134); and glioma and astrocytoma(NCT02924038).

Other checkpoint inhibitors that may be used in the present inventioninclude glucocorticoid-induced tumor necrosis factor receptor (GITR)agonists. GITR agonists that are being studied in clinical trialsinclude TRX518 (Leap Therapeutics), an agonistic anti-GITR antibody, inmalignant melanoma and other malignant solid tumors (NCT01239134 andNCT02628574); GWN323 (Novartis), an agonistic anti-GITR antibody, insolid tumors and lymphoma (NCT 02740270); INCAGN01876 (Incyte/Agenus),an agonistic anti-GITR antibody, in advanced cancers (NCT02697591 andNCT03126110); MK-4166 (Merck), an agonistic anti-GITR antibody, in solidtumors (NCT02132754) and MEDI1873 (Medimmune/AstraZeneca), an agonistichexameric GITR-ligand molecule with a human IgG1 Fc domain, in advancedsolid tumors (NCT02583165).

Other checkpoint inhibitors that may be used in the present inventioninclude inducible T-cell co-stimulator (ICOS, also known as CD278)agonists. ICOS agonists that are being studied in clinical trialsinclude MEDI-570 (Medimmune), an agonistic anti-ICOS antibody, inlymphomas (NCT02520791); GSK3359609 (Merck), an agonistic anti-ICOSantibody, in Phase 1 (NCT02723955); JTX-2011 (Jounce Therapeutics), anagonistic anti-ICOS antibody, in Phase 1 (NCT02904226).

Other checkpoint inhibitors that may be used in the present inventioninclude killer IgG-like receptor (KIR) inhibitors. KIR inhibitors thatare being studied in clinical trials include lirilumab(IPH2102/BMS-986015, Innate Pharma/Bristol-Myers Squibb), an anti-KIRantibody, in leukemias (NCT01687387, NCT02399917, NCT02481297,NCT02599649), multiple myeloma (NCT02252263), and lymphoma(NCT01592370); IPH2101 (1-7F9, Innate Pharma) in myeloma (NCT01222286and NCT01217203); and IPH4102 (Innate Pharma), an anti-KIR antibody thatbinds to three domains of the long cytoplasmic tail (KIR3DL2), inlymphoma (NCT02593045).

Other checkpoint inhibitors that may be used in the present inventioninclude CD47 inhibitors of interaction between CD47 and signalregulatory protein alpha (SIRPa). CD47/SIRPa inhibitors that are beingstudied in clinical trials include ALX-148 (Alexo Therapeutics), anantagonistic variant of (SIRPa) that binds to CD47 and preventsCD47/SIRPa-mediated signaling, in phase 1 (NCT03013218); TTI-621(SIRPa-Fc, Trillium Therapeutics), a soluble recombinant fusion proteincreated by linking the N-terminal CD47-binding domain of SIRPa with theFc domain of human IgG1, acts by binding human CD47, and preventing itfrom delivering its “do not eat” signal to macrophages, is in clinicaltrials in Phase 1 (NCT02890368 and NCT02663518); CC-90002 (Celgene), ananti-CD47 antibody, in leukemias (NCT02641002); and Hu5F9-G4 (FortySeven, Inc.), in colorectal neoplasms and solid tumors (NCT02953782),acute myeloid leukemia (NCT02678338) and lymphoma (NCT02953509).

Other checkpoint inhibitors that may be used in the present inventioninclude CD73 inhibitors. CD73 inhibitors that are being studied inclinical trials include MEDI9447 (Medimmune), an anti-CD73 antibody, insolid tumors (NCT02503774); and BMS-986179 (Bristol-Myers Squibb), ananti-CD73 antibody, in solid tumors (NCT02754141).

Other checkpoint inhibitors that may be used in the present inventioninclude agonists of stimulator of interferon genes protein (STING, alsoknown as transmembrane protein 173, or TMEM173). Agonists of STING thatare being studied in clinical trials include MK-1454 (Merck), anagonistic synthetic cyclic dinucleotide, in lymphoma (NCT03010176); andADU-S100 (MIW815, Aduro Biotech/Novartis), an agonistic synthetic cyclicdinucleotide, in Phase 1 (NCT02675439 and NCT03172936).

Other checkpoint inhibitors that may be used in the present inventioninclude CSF1R inhibitors. CSF1R inhibitors that are being studied inclinical trials include pexidartinib (PLX3397, Plexxikon), a CSF1R smallmolecule inhibitor, in colorectal cancer, pancreatic cancer, metastaticand advanced cancers (NCT02777710) and melanoma, non-small cell lungcancer, squamous cell head and neck cancer, gastrointestinal stromaltumor (GIST) and ovarian cancer (NCT02452424); and IMC-CS4 (LY3022855,Lilly), an anti-CSF-1R antibody, in pancreatic cancer (NCT03153410),melanoma (NCT03101254), and solid tumors (NCT02718911); and BLZ945(4-[2((1R,2R)-2-hydroxycyclohexylamino)-benzothiazol-6-yloxyl]-pyridine-2-carboxylicacid methylamide, Novartis), an orally available inhibitor of CSF1R, inadvanced solid tumors (NCT02829723).

Other checkpoint inhibitors that may be used in the present inventioninclude NKG2A receptor inhibitors. NKG2A receptor inhibitors that arebeing studied in clinical trials include monalizumab (IPH2201, InnatePharma), an anti-NKG2A antibody, in head and neck neoplasms(NCT02643550) and chronic lymphocytic leukemia (NCT02557516).

In some embodiments, the immune checkpoint inhibitor is selected fromnivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab,atezolizumab, or pidilizumab.

In another aspect, the present invention provides a method of treatingcancer in a patient in need thereof, wherein said method comprisesadministering to said patient a compound disclosed herein or apharmaceutically acceptable salt thereof in combination with one or moreadditional therapeutic agents selected from an indoleamine(2,3)-dioxygenase (IDO) inhibitor, a Poly ADP ribose polymerase (PARP)inhibitor, a histone deacetylase (HDAC) inhibitor, a CDK4/CDK6inhibitor, or a phosphatidylinositol 3 kinase (PI3K) inhibitor.

In some embodiments, the IDO inhibitor is selected from epacadostat,indoximod, capmanitib, GDC-0919, PF-06840003, BMS:F001287, Phy906/KD108,or an enzyme that breaks down kynurenine.

In some embodiments, the PARP inhibitor is selected from olaparib,rucaparib, or niraparib.

In some embodiments, the HDAC inhibitor is selected from vorinostat,romidepsin, panobinostat, belinostat, entinostat, or chidamide.

In some embodiments, the CDK 4/6 inhibitor is selected from palbociclib,ribociclib, abemaciclib or trilaciclib.

In some embodiments, the method further comprises administering to saidpatient a third therapeutic agent, such as an immune checkpointinhibitor. In some embodiments, the method comprises administering tothe patient in need thereof three therapeutic agents selected from acompound disclosed herein or a pharmaceutically acceptable salt thereof,a second therapeutic agent selected from an indoleamine(2,3)-dioxygenase (IDO) inhibitor, a Poly ADP ribose polymerase (PARP)inhibitor, a histone deacetylase (HDAC) inhibitor, a CDK4/CDK6inhibitor, or a phosphatidylinositol 3 kinase (PI3K) inhibitor, and athird therapeutic agent selected from an immune checkpoint inhibitor. Insome embodiments, the immune checkpoint inhibitor is selected fromnivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab,atezolizumab, or pidilizumab.

Another immunostimulatory therapeutic that may be used in the presentinvention is recombinant human interleukin 15 (rhIL-15). rhIL-15 hasbeen tested in the clinic as a therapy for melanoma and renal cellcarcinoma (NCT01021059 and NCT01369888) and leukemias (NCT02689453).Another immunostimulatory therapeutic that may be used in the presentinvention is recombinant human interleukin 12 (rhlL-12). Anothersuitable IL-15 based immunotherapeutic is heterodimeric IL-15 (hetlL-15,Novartis/Admune), a fusion complex composed of a synthetic form ofendogenous IL-15 complexed to the soluble IL-15 binding protein IL-15receptor alpha chain (IL15:sIL-15RA), which has been tested in Phase 1clinical trials for melanoma, renal cell carcinoma, non-small cell lungcancer and head and neck squamous cell carcinoma (NCT02452268).Recombinant human interleukin 12 (rhIL-12) has been tested in the clinicfor many oncological indications, for example, as a therapy for lymphoma(NM-IL-12, Neumedicines, Inc.), (NCT02544724 and NCT02542124).

In some embodiments, the PI3K inhibitor is selected from idelalisib,alpelisib, taselisib, pictilisib, copanlisib, duvelisib, PQR309, orTGR1202.

In another aspect, the present invention provides a method of treatingcancer in a patient in need thereof, wherein said method comprisesadministering to said patient a compound disclosed herein or apharmaceutically acceptable salt thereof in combination with one or moreadditional therapeutic agents selected from a platinum-basedtherapeutic, a taxane, a nucleoside inhibitor, or a therapeutic agentthat interferes with normal DNA synthesis, protein synthesis, cellreplication, or will otherwise inhibit rapidly proliferating cells.

In some embodiments, the platinum-based therapeutic is selected fromcisplatin, carboplatin, oxaliplatin, nedaplatin, picoplatin, orsatraplatin.

In some embodiments, the taxane is selected from paclitaxel, docetaxel,albumin-bound paclitaxel, cabazitaxel, or SID530.

In some embodiments, the therapeutic agent that interferes with normalDNA synthesis, protein synthesis, cell replication, or will otherwiseinterfere with the replication of rapidly proliferating cells isselected from trabectedin, mechlorethamine, vincristine, temozolomide,cytarabine, lomustine, azacitidine, omacetaxine mepesuccinate,asparaginase Erwinia chrysanthemi, eribulin mesylate, capacetrine,bendamustine, ixabepilone, nelarabine, clorafabine, trifluridine, ortipiracil.

In some embodiments, the method further comprises administering to saidpatient a third therapeutic agent, such as an immune checkpointinhibitor. In some embodiments, the method comprises administering tothe patient in need thereof three therapeutic agents selected from acompound disclosed herein or a pharmaceutically acceptable salt thereof,a second therapeutic agent selected from a platinum-based therapeutic, ataxane, a nucleoside inhibitor, or a therapeutic agent that interfereswith normal DNA synthesis, protein synthesis, cell replication, or willotherwise inhibit rapidly proliferating cells, and a third therapeuticagent selected from an immune checkpoint inhibitor.

In some embodiments, the immune checkpoint inhibitor is selected fromnivolumab, pembrolizumab, ipilimumab, avelumab, durvalumab,atezolizumab, or pidilizumab.

In some embodiments, any one of the foregoing methods further comprisesthe step of obtaining a biological sample from the patient and measuringthe amount of a disease-related biomarker.

In some embodiments, the biological sample is a blood sample.

In some embodiments, the disease-related biomarker is selected fromcirculating CD8+ T cells or the ratio of CD8+ T cells:Treg cells.

In one aspect, the present invention provides a method of treating anadvanced cancer, comprising administering a compound disclosed herein ora pharmaceutically acceptable salt thereof or pharmaceutical compositionthereof, either as a single agent (monotherapy), or in combination witha chemotherapeutic, a targeted therapeutic, such as a kinase inhibitor,and/or an immunomodulatory therapy, such as an immune checkpointinhibitor. In some embodiments, the immune checkpoint inhibitor is anantibody to PD-1. PD-1 binds to the programmed cell death 1 receptor(PD-1) to prevent the receptor from binding to the inhibitory ligandPDL-1, thus overriding the ability of tumors to suppress the hostanti-tumor immune response.

In some embodiments, the additional therapeutic agent is a kinaseinhibitor or VEGF-R antagonist. Approved VEGF inhibitors and kinaseinhibitors useful in the present invention include: bevacizumab(Avastin®, Genentech/Roche) an anti-VEGF monoclonal antibody;ramucirumab (Cyramza®, Eli Lilly), an anti-VEGFR-2 antibody andziv-aflibercept, also known as VEGF Trap (Zaltrap®; Regeneron/Sanofi).VEGFR inhibitors, such as regorafenib (Stivarga®, Bayer); vandetanib(Caprelsa®, AstraZeneca); axitinib (Inlyta®, Pfizer); and lenvatinib(Lenvima®, Eisai); Raf inhibitors, such as sorafenib (Nexavar®, Bayer AGand Onyx); dabrafenib (Tafinlar®, Novartis); and vemurafenib (Zelboraf®,Genentech/Roche); MEK inhibitors, such as cobimetanib (Cotellic®,Exelexis/Genentech/Roche); trametinib (Mekinist®, Novartis); Bcr-Abltyrosine kinase inhibitors, such as imatinib (Gleevec®, Novartis);nilotinib (Tasigna®, Novartis); dasatinib (Sprycel®,BristolMyersSquibb); bosutinib (Bosulif®, Pfizer); and ponatinib(Inclusig®, Ariad Pharmaceuticals); Her2 and EGFR inhibitors, such asgefitinib (Iressa®, AstraZeneca); erlotinib (Tarceeva®,Genentech/Roche/Astellas); lapatinib (Tykerb®, Novartis); afatinib(Gilotrif®, Boehringer Ingelheim); osimertinib (targeting activatedEGFR, Tagrisso®, AstraZeneca); and brigatinib (Alunbrig®, AriadPharmaceuticals); c-Met and VEGFR2 inhibitors, such as cabozanitib(Cometriq®, Exelexis); and multikinase inhibitors, such as sunitinib(Sutent®, Pfizer); pazopanib (Votrient®, Novartis); ALK inhibitors, suchas crizotinib (Xalkori®, Pfizer); ceritinib (Zykadia®, Novartis); andalectinib (Alecenza®, Genentech/Roche); Bruton's tyrosine kinaseinhibitors, such as ibrutinib (Imbruvica®, Pharmacyclics/Janssen); andFlt3 receptor inhibitors, such as midostaurin (Rydapt®, Novartis).

Other kinase inhibitors and VEGF-R antagonists that are in developmentand may be used in the present invention include tivozanib (AveoPharmaecuticals); vatalanib (Bayer/Novartis); lucitanib (ClovisOncology); dovitinib (TKI258, Novartis); Chiauanib (ChipscreenBiosciences); CEP-11981 (Cephalon); linifanib (Abbott Laboratories);neratinib (HKI-272, Puma Biotechnology); radotinib (Supect®, IY5511,Il-Yang Pharmaceuticals, S. Korea); ruxolitinib (Jakafi®, IncyteCorporation); PTC299 (PTC Therapeutics); CP-547,632 (Pfizer); foretinib(Exelexis, GlaxoSmithKline); quizartinib (Daiichi Sankyo) and motesanib(Amgen/Takeda).

In some embodiments, the additional therapeutic agent is an mTORinhibitor, which inhibits cell proliferation, angiogenesis and glucoseuptake. Approved mTOR inhibitors useful in the present invention includeeverolimus (Afinitor®, Novartis); temsirolimus (Torisel®, Pfizer); andsirolimus (Rapamune®, Pfizer).

In some embodiments, the additional therapeutic agent is a Poly ADPribose polymerase (PARP) inhibitor. Approved PARP inhibitors useful inthe present invention include olaparib (Lynparza®, AstraZeneca);rucaparib (Rubraca®, Clovis Oncology); and niraparib (Zejula®, Tesaro).Other PARP inhibitors being studied which may be used in the presentinvention include talazoparib (MDV3800/BMN 673/LT00673,Medivation/Pfizer/Biomarin); veliparib (ABT-888, AbbVie); and BGB-290(BeiGene, Inc.).

In some embodiments, the additional therapeutic agent is aphosphatidylinositol 3 kinase (PI3K) inhibitor. Approved PI3K inhibitorsuseful in the present invention include idelalisib (Zydelig®, Gilead).Other PI3K inhibitors being studied which may be used in the presentinvention include alpelisib (BYL719, Novartis); taselisib (GDC-0032,Genentech/Roche); pictilisib (GDC-0941, Genentech/Roche); copanlisib(BAY806946, Bayer); duvelisib (formerly IPI-145, InfinityPharmaceuticals); PQR309 (Piqur Therapeutics, Switzerland); and TGR1202(formerly RP5230, TG Therapeutics).

In some embodiments, the additional therapeutic agent is a proteasomeinhibitor. Approved proteasome inhibitors useful in the presentinvention include bortezomib (Velcade®, Takeda); carfilzomib (Kyprolis®,Amgen); and ixazomib (Ninlaro®, Takeda).

In some embodiments, the additional therapeutic agent is a histonedeacetylase (HDAC) inhibitor. Approved HDAC inhibitors useful in thepresent invention include vorinostat (Zolinza®, Merck); romidepsin(Istodax®, Celgene); panobinostat (Farydak®, Novartis); and belinostat(Beleodaq®, Spectrum Pharmaceuticals). Other HDAC inhibitors beingstudied which may be used in the present invention include entinostat(SNDX-275, Syndax Pharmaceuticals) (NCT00866333); and chidamide(Epidaza®, HBI-8000, Chipscreen Biosciences, China).

In some embodiments, the additional therapeutic agent is a CDKinhibitor, such as a CDK 4/6 inhibitor. Approved CDK 4/6 inhibitorsuseful in the present invention include palbociclib (Ibrance®, Pfizer);and ribociclib (Kisqali®, Novartis). Other CDK 4/6 inhibitors beingstudied which may be used in the present invention include abemaciclib(Ly2835219, Eli Lilly); and trilaciclib (G1T28, G1 Therapeutics).

In some embodiments, the additional therapeutic agent is an indoleamine(2,3)-dioxygenase (IDO) inhibitor. IDO inhibitors being studied whichmay be used in the present invention include epacadostat (INCB024360,Incyte); indoximod (NLG-8189, NewLink Genetics Corporation); capmanitib(INC280, Novartis); GDC-0919 (Genentech/Roche); PF-06840003 (Pfizer);BMS:F001287 (Bristol-Myers Squibb); Phy906/KD108 (Phytoceutica); and anenzyme that breaks down kynurenine (Kynase, Kyn Therapeutics).

In some embodiments, the additional therapeutic agent is a growth factorantagonist, such as an antagonist of platelet-derived growth factor(PDGF), or epidermal growth factor (EGF) or its receptor (EGFR).Approved PDGF antagonists which may be used in the present inventioninclude olaratumab (Lartruvo®; Eli Lilly). Approved EGFR antagonistswhich may be used in the present invention include cetuximab (Erbitux®,Eli Lilly); necitumumab (Portrazza®, Eli Lilly), panitumumab (Vectibix®,Amgen); and osimertinib (targeting activated EGFR, Tagrisso®,AstraZeneca).

In some embodiments, the additional therapeutic agent is an aromataseinhibitor. Approved aromatase inhibitors which may be used in thepresent invention include exemestane (Aromasin®, Pfizer); anastazole(Arimidex®, AstraZeneca) and letrozole (Femora®, Novartis).

In some embodiments, the additional therapeutic agent is an antagonistof the hedgehog pathway. Approved hedgehog pathway inhibitors which maybe used in the present invention include sonidegib (Odomzo®, SunPharmaceuticals); and vismodegib (Erivedge®, Genentech), both fortreatment of basal cell carcinoma.

In some embodiments, the additional therapeutic agent is a folic acidinhibitor. Approved folic acid inhibitors useful in the presentinvention include pemetrexed (Alimta®, Eli Lilly).

In some embodiments, the additional therapeutic agent is a CC chemokinereceptor 4 (CCR4) inhibitor. CCR4 inhibitors being studied that may beuseful in the present invention include mogamulizumab (Poteligeo®, KyowaHakko Kirin, Japan).

In some embodiments, the additional therapeutic agent is an isocitratedehydrogenase (IDH) inhibitor. IDH inhibitors being studied which may beused in the present invention include AG120 (Celgene; NCT02677922);AG221 (Celgene, NCT02677922; NCT02577406); BAY1436032 (Bayer,NCT02746081); IDH305 (Novartis, NCT02987010).

In some embodiments, the additional therapeutic agent is an arginaseinhibitor. Arginase inhibitors being studied which may be used in thepresent invention include AEB1102 (pegylated recombinant arginase,Aeglea Biotherapeutics), which is being studied in Phase 1 clinicaltrials for acute myeloid leukemia and myelodysplastic syndrome(NCT02732184) and solid tumors (NCT02561234); and CB-1158 (CalitheraBiosciences).

In some embodiments, the additional therapeutic agent is a glutaminaseinhibitor. Glutaminase inhibitors being studied which may be used in thepresent invention include CB-839 (Calithera Biosciences).

In some embodiments, the additional therapeutic agent is an antibodythat binds to tumor antigens, that is, proteins expressed on the cellsurface of tumor cells. Approved antibodies that bind to tumor antigenswhich may be used in the present invention include rituximab (Rituxan®,Genentech/BiogenIdec); ofatumumab (anti-CD20, Arzerra®,GlaxoSmithKline); obinutuzumab (anti-CD20, Gazyva®, Genentech),ibritumomab (anti-CD20 and Yttrium-90, Zevalin®, SpectrumPharmaceuticals); daratumumab (anti-CD38, Darzalex®, Janssen Biotech),dinutuximab (anti-glycolipid GD2, Unituxin®, United Therapeutics);trastuzumab (anti-HER2, Herceptin®, Genentech); ado-trastuzumabemtansine (anti-HER2, fused to emtansine, Kadcyla®, Genentech); andpertuzumab (anti-HER2, Perjeta®, Genentech); and brentuximab vedotin(anti-CD30-drug conjugate, Adcetris®, Seattle Genetics).

In some embodiments, the additional therapeutic agent is a topoisomeraseinhibitor. Approved topoisomerase inhibitors useful in the presentinvention include irinotecan (Onivyde®, Merrimack Pharmaceuticals);topotecan (Hycamtin®, GlaxoSmithKline). Topoisomerase inhibitors beingstudied which may be used in the present invention include pixantrone(Pixuvri®, CTI Biopharma).

In some embodiments, the additional therapeutic agent is a nucleosideinhibitor, or other therapeutic that interfere with normal DNAsynthesis, protein synthesis, cell replication, or will otherwiseinhibit rapidly proliferating cells. Such nucleoside inhibitors or othertherapeutics include trabectedin (guanidine alkylating agent, Yondelis®,Janssen Oncology), mechlorethamine (alkylating agent, Valchlor®,Aktelion Pharmaceuticals); vincristine (Oncovin®, Eli Lilly; Vincasar®,Teva Pharmaceuticals; Marqibo®, Talon Therapeutics); temozolomide(prodrug to alkylating agent5-(3-methyltriazen-1-yl)-imidazole-4-carboxamide (MTIC) Temodar®,Merck); cytarabine injection (ara-C, antimetabolic cytidine analog,Pfizer); lomustine (alkylating agent, CeeNU®, Bristol-Myers Squibb;Gleostine®, NextSource Biotechnology); azacitidine (pyrimidinenucleoside analog of cytidine, Vidaza®, Celgene); omacetaxinemepesuccinate (cephalotaxine ester) (protein synthesis inhibitor,Synribo®; Teva Pharmaceuticals); asparaginase Erwinia chrysanthemi(enzyme for depletion of asparagine, Elspar®, Lundbeck; Erwinaze®, EUSAPharma); eribulin mesylate (microtubule inhibitor, tubulin-basedantimitotic, Halaven®, Eisai); cabazitaxel (microtubule inhibitor,tubulin-based antimitotic, Jevtana®, Sanofi-Aventis); capacetrine(thymidylate synthase inhibitor, Xeloda®, Genentech); bendamustine(bifunctional mechlorethamine derivative, believed to form interstrandDNA cross-links, Treanda®, Cephalon/Teva); ixabepilone (semi-syntheticanalog of epothilone B, microtubule inhibitor, tubulin-basedantimitotic, Ixempra®, Bristol-Myers Squibb); nelarabine (prodrug ofdeoxyguanosine analog, nucleoside metabolic inhibitor, Arranon®,Novartis); clorafabine (prodrug of ribonucleotide reductase inhibitor,competitive inhibitor of deoxycytidine, Clolar®, Sanofi-Aventis); andtrifluridine and tipiracil (thymidine-based nucleoside analog andthymidine phosphorylase inhibitor, Lonsurf®, Taiho Oncology).

In some embodiments, the additional therapeutic agent is aplatinum-based therapeutic, also referred to as platins. Platins causecross-linking of DNA, such that they inhibit DNA repair and/or DNAsynthesis, mostly in rapidly reproducing cells, such as cancer cells.Approved platinum-based therapeutics which may be used in the presentinvention include cisplatin (Platinol®, Bristol-Myers Squibb);carboplatin (Paraplatin®, Bristol-Myers Squibb; also, Teva; Pfizer);oxaliplatin (Eloxitin® Sanofi-Aventis); and nedaplatin (Aqupla®,Shionogi). Other platinum-based therapeutics which have undergoneclinical testing and may be used in the present invention includepicoplatin (Poniard Pharmaceuticals); and satraplatin (JM-216, Agennix).

In some embodiments, the additional therapeutic agent is a taxanecompound, which causes disruption of microtubules, which are essentialfor cell division. Approved taxane compounds which may be used in thepresent invention include paclitaxel (Taxol®, Bristol-Myers Squibb),docetaxel (Taxotere®, Sanofi-Aventis; Docefrez®, Sun Pharmaceutical),albumin-bound paclitaxel (Abraxane®; Abraxis/Celgene), and cabazitaxel(Jevtana®, Sanofi-Aventis). Other taxane compounds which have undergoneclinical testing and may be used in the present invention include SID530(SK Chemicals, Co.) (NCT00931008).

In some embodiments, the additional therapeutic agent is an inhibitor ofanti-apoptotic proteins, such as BCL-2. Approved anti-apoptotics whichmay be used in the present invention include venetoclax (Venclexta®,AbbVie/Genentech); and blinatumomab (Blincyto®, Amgen). Othertherapeutic agents targeting apoptotic proteins which have undergoneclinical testing and may be used in the present invention includenavitoclax (ABT-263, Abbott), a BCL-2 inhibitor (NCT02079740).

In some embodiments, the present invention provides a method of treatingprostate cancer comprising administering to a patient in need thereof aneffective amount of a compound disclosed herein or a pharmaceuticallyacceptable salt thereof or pharmaceutical composition thereof incombination with an additional therapeutic agent that interferes withthe synthesis or activity of androgens. Approved androgen receptorinhibitors useful in the present invention include enzalutamide(Xtandi®, Astellas/Medivation); approved inhibitors of androgensynthesis include abiraterone (Zytiga®, Centocor/Ortho); approvedantagonist of gonadotropin-releasing hormone (GnRH) receptor (degaralix,Firmagon®, Ferring Pharmaceuticals).

In some embodiments, the additional therapeutic agent is a selectiveestrogen receptor modulator (SERM), which interferes with the synthesisor activity of estrogens. Approved SERMs useful in the present inventioninclude raloxifene (Evista®, Eli Lilly).

In some embodiments, the additional therapeutic agent is an inhibitor ofbone resorption. An approved therapeutic which inhibits bone resorptionis Denosumab (Xgeva®, Amgen), an antibody that binds to RANKL, preventsbinding to its receptor RANK, found on the surface of osteoclasts, theirprecursors, and osteoclast-like giant cells, which mediates bonepathology in solid tumors with osseous metastases. Other approvedtherapeutics that inhibit bone resorption include bisphosphonates, suchas zoledronic acid (Zometa®, Novartis).

In some embodiments, the additional therapeutic agent is an inhibitor ofinteraction between the two primary p53 suppressor proteins, MDMX andMDM2. Inhibitors of p53 suppression proteins being studied which may beused in the present invention include ALRN-6924 (Aileron), a stapledpeptide that equipotently binds to and disrupts the interaction of MDMXand MDM2 with p53. ALRN-6924 is currently being evaluated in clinicaltrials for the treatment of AML, advanced myelodysplastic syndrome (MDS)and peripheral T-cell lymphoma (PTCL) (NCT02909972; NCT02264613).

In some embodiments, the additional therapeutic agent is an inhibitor oftransforming growth factor-beta (TGF-beta or TGFβ). Inhibitors ofTGF-beta proteins being studied which may be used in the presentinvention include NIS793 (Novartis), an anti-TGF-beta antibody beingtested in the clinic for treatment of various cancers, including breast,lung, hepatocellular, colorectal, pancreatic, prostate and renal cancer(NCT 02947165). In some embodiments, the inhibitor of TGF-beta proteinsis fresolimumab (GC1008; Sanofi-Genzyme), which is being studied formelanoma (NCT00923169); renal cell carcinoma (NCT00356460); andnon-small cell lung cancer (NCT02581787). Additionally, in someembodiments, the additional therapeutic agent is a TGF-beta trap, suchas described in Connolly et al. (2012) Int'l J. Biological Sciences8:964-978. One therapeutic compound currently in clinical trials fortreatment of solid tumors is M7824 (Merck KgaA—formerly MSB0011459X),which is a bispecific, anti-PD-L1/TGFβ trap compound (NCT02699515); and(NCT02517398). M7824 is comprised of a fully human IgG1 antibody againstPD-L1 fused to the extracellular domain of human TGF-beta receptor II,which functions as a TGFβ “trap.”

Additional Co Administered Therapeutic Agents -Targeted Therapeutics andImmunomodulatory Drugs

In some embodiments, the additional therapeutic agent is selected from atargeted therapeutic or immunomodulatory drug. Adjuvant therapies withtargeted therapeutics or immunomodulatory drugs have shown promisingeffectiveness when administered alone but are limited by the developmentof tumor immunity over time or evasion of the immune response.

In some embodiments, the present invention provides a method of treatingcancer, such as a cancer described herein, comprising administering to apatient in need thereof an effective amount of a compound disclosedherein or a pharmaceutically acceptable salt thereof or pharmaceuticalcomposition thereof in combination with an additional therapeutic agentsuch as a targeted therapeutic or an immunomodulatory drug. In someembodiments, the immunomodulatory therapeutic specifically inducesapoptosis of tumor cells. Approved immunomodulatory therapeutics whichmay be used in the present invention include pomalidomide (Pomalyst®,Celgene); lenalidomide (Revlimid®, Celgene); ingenol mebutate (Picato®,LEO Pharma).

In other embodiments, the immunomodulatory therapeutic is a cancervaccine. In some embodiments, the cancer vaccine is selected fromsipuleucel-T (Provenge®, Dendreon/Valeant Pharmaceuticals), which hasbeen approved for treatment of asymptomatic, or minimally symptomaticmetastatic castrate-resistant (hormone-refractory) prostate cancer; andtalimogene laherparepvec (Imlygic®, BioVex/Amgen, previously known asT-VEC), a genetically modified oncolytic viral therapy approved fortreatment of unresectable cutaneous, subcutaneous and nodal lesions inmelanoma. In some embodiments, the additional therapeutic agent isselected from an oncolytic viral therapy such as pexastimogenedevacirepvec (PexaVec/JX-594, SillaJen/formerly JennerexBiotherapeutics), a thymidine kinase- (TK-) deficient vaccinia virusengineered to express GM-CSF, for hepatocellular carcinoma (NCT02562755)and melanoma (NCT00429312); pelareorep (Reolysin®, Oncolytics Biotech),a variant of respiratory enteric orphan virus (reovirus) which does notreplicate in cells that are not RAS-activated, in numerous cancers,including colorectal cancer (NCT01622543); prostate cancer(NCT01619813); head and neck squamous cell cancer (NCT01166542);pancreatic adenocarcinoma (NCT00998322); and non-small cell lung cancer(NSCLC) (NCT 00861627); enadenotucirev (NG-348, PsiOxus, formerly knownas ColoAd1), an adenovirus engineered to express a full length CD80 andan antibody fragment specific for the T-cell receptor CD3 protein, inovarian cancer (NCT02028117); metastatic or advanced epithelial tumorssuch as in colorectal cancer, bladder cancer, head and neck squamouscell carcinoma and salivary gland cancer (NCT02636036); ONCOS-102(Targovax/formerly Oncos), an adenovirus engineered to express GM-CSF,in melanoma (NCT03003676); and peritoneal disease, colorectal cancer orovarian cancer (NCT02963831); GL-ONC1 (GLV-1h68/GLV-1h153, GeneluxGmbH), vaccinia viruses engineered to express beta-galactosidase(beta-gal)/beta-glucoronidase or beta-gal/human sodium iodide symporter(hNIS), respectively, were studied in peritoneal carcinomatosis(NCT01443260); fallopian tube cancer, ovarian cancer (NCT 02759588); orCG0070 (Cold Genesys), an adenovirus engineered to express GM-CSF, inbladder cancer (NCT02365818).

In some embodiments, the additional therapeutic agent is selected fromJX-929 (SillaJen/formerly Jennerex Biotherapeutics), a TK- and vacciniagrowth factor-deficient vaccinia virus engineered to express cytosinedeaminase, which is able to convert the prodrug 5-fluorocytosine to thecytotoxic drug 5-fluorouracil; TG01 and TGO2 (Targovax/formerly Oncos),peptide-based immunotherapy agents targeted for difficult-to-treat RASmutations; and TILT-123 (TILT Biotherapeutics), an engineered adenovirusdesignated: Ad5/3-E2F-delta24-hTNFa-IRES-hIL20; and VSV-GP(ViraTherapeutics) a vesicular stomatitis virus (VSV) engineered toexpress the glycoprotein (GP) of lymphocytic choriomeningitis virus(LCMV), which can be further engineered to express antigens designed toraise an antigen-specific CD8⁺ T cell response.

In some embodiments, the present invention comprises administering tosaid patient a compound disclosed herein or a pharmaceuticallyacceptable salt thereof in combination with a T-cell engineered toexpress a chimeric antigen receptor, or CAR. The T-cells engineered toexpress such chimeric antigen receptor are referred to as a CAR-T cells.

CARs have been constructed that consist of binding domains, which may bederived from natural ligands, single chain variable fragments (scFv)derived from monoclonal antibodies specific for cell-surface antigens,fused to endodomains that are the functional end of the T-cell receptor(TCR), such as the CD3-zeta signaling domain from TCRs, which is capableof generating an activation signal in T lymphocytes. Upon antigenbinding, such CARs link to endogenous signaling pathways in the effectorcell and generate activating signals similar to those initiated by theTCR complex.

For example, in some embodiments the CAR-T cell is one of thosedescribed in U.S. Pat. No. 8,906,682 (June; hereby incorporated byreference in its entirety), which discloses CAR-T cells engineered tocomprise an extracellular domain having an antigen binding domain (suchas a domain that binds to CD19), fused to an intracellular signalingdomain of the T cell antigen receptor complex zeta chain (such as CD3zeta). When expressed in the T cell, the CAR is able to redirect antigenrecognition based on the antigen binding specificity. In the case ofCD19, the antigen is expressed on malignant B cells. Over 200 clinicaltrials are currently in progress employing CAR-T in a wide range ofindications.[https://clinicaltrials.gov/ct2/results?term=chimeric+antigen+receptors&pg=1].

Additional Co-Administered Therapeutic Agents—Immunostimulatory Drugs

In some embodiments, the additional therapeutic agent is animmunostimulatory drug. For example, antibodies blocking the PD-1 andPD-L1 inhibitory axis can unleash activated tumor-reactive T cells andhave been shown in clinical trials to induce durable anti-tumorresponses in increasing numbers of tumor histologies, including sometumor types that conventionally have not been considered immunotherapysensitive. See, e.g., Okazaki, T. et al. (2013) Nat. Immunol. 14,1212-1218; Zou et al. (2016) Sci. Transl. Med. 8. The anti-PD-1 antibodynivolumab (Opdivo®, Bristol-Myers Squibb, also known as ONO-4538,MDX1106 and BMS-936558), has shown potential to improve the overallsurvival in patients with RCC who had experienced disease progressionduring or after prior anti-angiogenic therapy.

In some embodiments, the present invention provides a method of treatingcancer, such as a cancer described herein, comprising administering to apatient in need thereof an effective amount of a compound disclosedherein or a pharmaceutically acceptable salt thereof or pharmaceuticalcomposition thereof in combination with an additional therapeutic agentsuch as a immunostimulatory drug, such as an immune checkpointinhibitor. In some embodiments, the compound and the checkpointinhibitor are administered simultaneously or sequentially. In someembodiments, a compound disclosed herein is administered prior to theinitial dosing with the immune checkpoint inhibitor. In certainembodiments, the immune checkpoint inhibitor is administered prior tothe initial dosing with the compound disclosed herein.

In certain embodiments, the immune checkpoint inhibitor is selected froma PD-1 antagonist, a PD-L1 antagonist, or a CTLA-4 antagonist. In someembodiments, a CXCR4 antagonist such as a compound disclosed herein or apharmaceutically acceptable salt thereof is administered in combinationwith nivolumab (anti-PD-1 antibody, Opdivo®, Bristol-Myers Squibb);pembrolizumab (anti-PD-1 antibody, Keytruda®, Merck); ipilimumab(anti-CTLA-4 antibody, Yervoy®, Bristol-Myers Squibb); durvalumab(anti-PD-L1 antibody, Imfinzi®, AstraZeneca); or atezolizumab(anti-PD-L1 antibody, Tecentriq®, Genentech).

Other immune checkpoint inhibitors suitable for use in the presentinvention include REGN2810 (Regeneron), an anti-PD-1 antibody tested inpatients with basal cell carcinoma (NCT03132636); NSCLC (NCT03088540);cutaneous squamous cell carcinoma (NCT02760498); lymphoma (NCT02651662);and melanoma (NCT03002376); pidilizumab (CureTech), also known asCT-011, an antibody that binds to PD-1, in clinical trials for diffuselarge B-cell lymphoma and multiple myeloma; avelumab (Bavencio®,Pfizer/Merck KGaA), also known as MSB0010718C), a fully human IgG1anti-PD-L1 antibody, in clinical trials for non-small cell lung cancer,Merkel cell carcinoma, mesothelioma, solid tumors, renal cancer, ovariancancer, bladder cancer, head and neck cancer, and gastric cancer; andPDR001 (Novartis), an inhibitory antibody that binds to PD-1, inclinical trials for non-small cell lung cancer, melanoma, triplenegative breast cancer and advanced or metastatic solid tumors.Tremelimumab (CP-675,206; Astrazeneca) is a fully human monoclonalantibody against CTLA-4 that has been in studied in clinical trials fora number of indications, including: mesothelioma, colorectal cancer,kidney cancer, breast cancer, lung cancer and non-small cell lungcancer, pancreatic ductal adenocarcinoma, pancreatic cancer, germ cellcancer, squamous cell cancer of the head and neck, hepatocellularcarcinoma, prostate cancer, endometrial cancer, metastatic cancer in theliver, liver cancer, large B-cell lymphoma, ovarian cancer, cervicalcancer, metastatic anaplastic thyroid cancer, urothelial cancer,fallopian tube cancer, multiple myeloma, bladder cancer, soft tissuesarcoma, and melanoma. AGEN-1884 (Agenus) is an anti-CTLA4 antibody thatis being studied in Phase 1 clinical trials for advanced solid tumors(NCT02694822).

Another paradigm for immune-stimulation is the use of oncolytic viruses.In some embodiments, the present invention provides a method fortreating a patient by administering a CXCR4 antagonist such as acompound disclosed herein or a pharmaceutically acceptable salt thereofor pharmaceutical composition thereof in combination with animmunostimulatory therapy such as oncolytic viruses. Approvedimmunostimulatory oncolytic viruses which may be used in the presentinvention include talimogene laherparepvec (live, attenuated herpessimplex virus, Imlygic®, Amgen).

In some embodiments, the additional therapeutic agent is an activator ofretinoic acid receptor-related orphan receptor γ (RORyt). RORyt is atranscription factor with key roles in the differentiation andmaintenance of Type 17 effector subsets of CD4+(Th17) and CD8+(Tc17) Tcells, as well as the differentiation of IL-17 expressing innate immunecell subpopulations such as NK cells. An activator of RORyt, that isbeing studied which may be used in the present invention is LYC-55716(Lycera), which is currently being evaluated in clinical trials for thetreatment of solid tumors (NCT02929862).

In some embodiments, the additional therapeutic agent is an agonist oractivator of a toll-like receptor (TLR). Suitable activators of TLRsinclude an agonist or activator of TLR9 such as SD-101 (Dynavax). SD-101is an immunostimulatory CpG which is being studied for B-cell,follicular and other lymphomas (NCT02254772). Agonists or activators ofTLR8 which may be used in the present invention include motolimod(VTX-2337, VentiRx Pharmaceuticals) which is being studied for squamouscell cancer of the head and neck (NCT02124850) and ovarian cancer(NCT02431559).

Other checkpoint inhibitors that may be used in the present inventioninclude inhibitors of T-cell immunoglobulin mucin containing protein-3(TIM-3). TIM-3 inhibitors that may be used in the present inventioninclude TSR-022, LY3321367 and MBG453. TSR-022 (Tesaro) is an anti-TIM-3antibody which is being studied in solid tumors (NCT02817633). LY3321367(Eli Lilly) is an anti-TIM-3 antibody which is being studied in solidtumors (NCT03099109). MBG453 (Novartis) is an anti-TIM-3 antibody whichis being studied in advanced malignancies (NCT02608268).

Other checkpoint inhibitors that may be used in the present inventioninclude inhibitors of T cell immunoreceptor with Ig and ITIM domains, orTIGIT, an immune receptor on certain T cells and NK cells. TIGITinhibitors that may be used in the present invention include BMS-986207(Bristol-Myers Squibb), an anti-TIGIT monoclonal antibody (NCT02913313);OMP-313M32 (Oncomed); and anti-TIGIT monoclonal antibody (NCT03119428).

Checkpoint inhibitors that may be used in the present invention alsoinclude inhibitors of Lymphocyte Activation Gene-3 (LAG-3). LAG-3inhibitors that may be used in the present invention include BMS-986016and REGN3767 and IMP321. BMS-986016 (Bristol-Myers Squibb), ananti-LAG-3 antibody, is being studied in glioblastoma and gliosarcoma(NCT02658981). REGN3767 (Regeneron), is also an anti-LAG-3 antibody, andis being studied in malignancies (NCT03005782). IMP321 (Immutep S.A.) isan LAG-3-Ig fusion protein, being studied in melanoma (NCT02676869);adenocarcinoma (NCT02614833); and metastatic breast cancer(NCT00349934).

Other immune-oncology agents that may be used in the present inventionin combination with CXCR4 inhibitors such as a compound disclosed hereininclude urelumab (BMS-663513, Bristol-Myers Squibb), an anti-CD137monoclonal antibody; varlilumab (CDX-1127, Celldex Therapeutics), ananti-CD27 monoclonal antibody; BMS-986178 (Bristol-Myers Squibb), ananti-OX40 monoclonal antibody; lirilumab (IPH2102/BMS-986015, InnatePharma, Bristol-Myers Squibb), an anti-KIR monoclonal antibody;monalizumab (IPH2201, Innate Pharma, AstraZeneca) an anti-NKG2Amonoclonal antibody; andecaliximab (GS-5745, Gilead Sciences), ananti-MMP9 antibody; MK-4166 (Merck & Co.), an anti-GITR monoclonalantibody.

Other additional therapeutic agents that may be used in the presentinvention include glembatumumab vedotin-monomethyl auristatin E (MMAE)(Celldex), an anti-glycoprotein NMB (gpNMB) antibody (CR011) linked tothe cytotoxic MMAE. gpNMB is a protein overexpressed by multiple tumortypes associated with cancer cells' ability to metastasize.

A compound of the current invention may also be used to advantage incombination with other antiproliferative compounds. Suchantiproliferative compounds include, but are not limited to checkpointinhibitors; aromatase inhibitors; antiestrogens; topoisomerase Iinhibitors; topoisomerase II inhibitors; microtubule active compounds;alkylating compounds; histone deacetylase inhibitors; compounds whichinduce cell differentiation processes; cyclooxygenase inhibitors; MMPinhibitors; mTOR inhibitors; antineoplastic antimetabolites; platincompounds; compounds targeting/decreasing a protein or lipid kinaseactivity and further anti-angiogenic compounds; compounds which target,decrease or inhibit the activity of a protein or lipid phosphatase;gonadorelin agonists; anti-androgens; methionine aminopeptidaseinhibitors; matrix metalloproteinase inhibitors; bisphosphonates;biological response modifiers; antiproliferative antibodies; heparanaseinhibitors; inhibitors of Ras oncogenic isoforms; telomerase inhibitors;proteasome inhibitors; compounds used in the treatment of hematologicmalignancies; compounds which target, decrease or inhibit the activityof Flt-3; Hsp90 inhibitors such as 17-AAG (17-allylaminogeldanamycin,NSC330507), 17-DMAG(17-dimethylaminoethylamino-17-demethoxy-geldanamycin, NSC707545),IPI-504, CNF1010, CNF2024, CNF1010 from Conforma Therapeutics;temozolomide (Temodal); kinesin spindle protein inhibitors, such asSB715992 or SB743921 from GlaxoSmithKline, or pentamidine/chlorpromazinefrom CombinatoRx; MEK inhibitors such as ARRY142886 from ArrayBioPharma, AZd₆244 from AstraZeneca, PD181461 from Pfizer andleucovorin.

The term “checkpoint inhibitor” as used herein relates to agents usefulin preventing cancer cells from avoiding the immune system of thepatient. One of the major mechanisms of anti-tumor immunity subversionis known as “T-cell exhaustion,” which results from chronic exposure toantigens that has led to up-regulation of inhibitory receptors. Theseinhibitory receptors serve as immune checkpoints in order to preventuncontrolled immune reactions.

PD-1 and co-inhibitory receptors such as cytotoxic T-lymphocyte antigen4 (CTLA-4, B and T Lymphocyte Attenuator (BTLA; CD272), T cellImmunoglobulin and Mucin domain-3 (Tim-3), Lymphocyte Activation Gene-3(Lag-3; CD223), and others are often referred to as a checkpointregulators. They act as molecular “gatekeepers” that allow extracellularinformation to dictate whether cell cycle progression and otherintracellular signalling processes should proceed.

In one aspect, the checkpoint inhibitor is a biologic therapeutic or asmall molecule. In another aspect, the checkpoint inhibitor is amonoclonal antibody, a humanized antibody, a fully human antibody, afusion protein or a combination thereof. In a further aspect, thecheckpoint inhibitor inhibits a checkpoint protein selected from CTLA-4,PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR,2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligands or acombination thereof. In an additional aspect, the checkpoint inhibitorinteracts with a ligand of a checkpoint protein selected from CTLA-4,PDL1, PDL2, PD1, B7-H3, B7-H4, BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR,2B4, CD160, CGEN-15049, CHK 1, CHK2, A2aR, B-7 family ligands or acombination thereof. In an aspect, the checkpoint inhibitor is animmunostimulatory agent, a T cell growth factor, an interleukin, anantibody, a vaccine or a combination thereof. In a further aspect, theinterleukin is IL-7 or IL-15. In a specific aspect, the interleukin isglycosylated IL-7. In an additional aspect, the vaccine is a dendriticcell (DC) vaccine.

Checkpoint inhibitors include any agent that blocks or inhibits in astatistically significant manner, the inhibitory pathways of the immunesystem. Such inhibitors may include small molecule inhibitors or mayinclude antibodies, or antigen binding fragments thereof, that bind toand block or inhibit immune checkpoint receptors or antibodies that bindto and block or inhibit immune checkpoint receptor ligands. Illustrativecheckpoint molecules that may be targeted for blocking or inhibitioninclude, but are not limited to, CTLA-4, PDL1, PDL2, PD1, B7-H3, B7-H4,BTLA, HVEM, GAL9, LAG3, TIM3, VISTA, KIR, 2B4 (belongs to the CD2 familyof molecules and is expressed on all NK, γδ, and memory CD8⁺ (αβ) Tcells), CD160 (also referred to as BY55), CGEN-15049, CHK 1 and CHK2kinases, A2aR, and various B-7 family ligands. B7 family ligandsinclude, but are not limited to, B7- 1, B7-2, B7-DC, B7-H1, B7-H2,B7-H3, B7-H4, B7-H5, B7-H6 and B7-H7. Checkpoint inhibitors includeantibodies, or antigen binding fragments thereof, other bindingproteins, biologic therapeutics, or small molecules, that bind to andblock or inhibit the activity of one or more of CTLA-4, PDL1, PDL2, PD1,BTLA, HVEM, TIM3, GAL9, LAG3, VISTA, KIR, 2B4, CD 160 and CGEN-15049.Illustrative immune checkpoint inhibitors include Tremelimumab (CTLA-4blocking antibody), anti-OX40, PD-L1 monoclonal Antibody (Anti-B7-H1;MEDI4736), MK-3475 (PD-1 blocker), Nivolumab (anti-PD1 antibody), CT-011(anti-PD1 antibody), BY55 monoclonal antibody, AMP224 (anti-PDL1antibody), BMS- 936559 (anti-PDL1 antibody), MPLDL3280A (anti-PDL1antibody), MSB0010718C (anti-PDL1 antibody), and ipilimumab (anti-CTLA-4checkpoint inhibitor). Checkpoint protein ligands include, but are notlimited to PD-L1, PD-L2, B7-H3, B7-H4, CD28, CD86 and TIM-3.

In certain embodiments, the immune checkpoint inhibitor is selected froma PD-1 antagonist, a PD-L1 antagonist, and a CTLA-4 antagonist. In someembodiments, the checkpoint inhibitor is selected from the groupconsisting of nivolumab (Opdivo®), ipilimumab (Yervoy®), andpembrolizumab (Keytruda®).

In some embodiments, the checkpoint inhibitor is selected from the groupconsisting of lambrolizumab (MK-3475), nivolumab (BMS-936558),pidilizumab (CT-011), AMP-224, MDX-1105, MEDI4736, MPDL3280A,BMS-936559, ipilimumab, lirlumab, IPH2101, pembrolizumab (Keytruda®),and tremelimumab.

The term “aromatase inhibitor” as used herein relates to a compoundwhich inhibits estrogen production, for instance, the conversion of thesubstrates androstenedione and testosterone to estrone and estradiol,respectively. The term includes, but is not limited to steroids,especially atamestane, exemestane and formestane and, in particular,non-steroids, especially aminoglutethimide, roglethimide,pyridoglutethimide, trilostane, testolactone, ketokonazole, vorozole,fadrozole, anastrozole and letrozole. Exemestane is marketed under thetrade name Aromasin™. Formestane is marketed under the trade nameLentaron™. Fadrozole is marketed under the trade name Afema™.Anastrozole is marketed under the trade name Arimidex™. Letrozole ismarketed under the trade names Femara™ or Femar™ Aminoglutethimide ismarketed under the trade name Orimeten™. A combination of the inventioncomprising a chemotherapeutic agent which is an aromatase inhibitor isparticularly useful for the treatment of hormone receptor positivetumors, such as breast tumors.

The term “antiestrogen” as used herein relates to a compound whichantagonizes the effect of estrogens at the estrogen receptor level. Theterm includes, but is not limited to tamoxifen, fulvestrant, raloxifeneand raloxifene hydrochloride. Tamoxifen is marketed under the trade nameNolvadex™. Raloxifene hydrochloride is marketed under the trade nameEvista™. Fulvestrant can be administered under the trade name Faslodex™.A combination of the invention comprising a chemotherapeutic agent whichis an antiestrogen is particularly useful for the treatment of estrogenreceptor positive tumors, such as breast tumors.

The term “anti-androgen” as used herein relates to any substance whichis capable of inhibiting the biological effects of androgenic hormonesand includes, but is not limited to, bicalutamide (Casodex™). The term“gonadorelin agonist” as used herein includes, but is not limited toabarelix, goserelin and goserelin acetate. Goserelin can be administeredunder the trade name Zoladex™.

The term “topoisomerase I inhibitor” as used herein includes, but is notlimited to topotecan, gimatecan, irinotecan, camptothecian and itsanalogues, 9-nitrocamptothecin and the macromolecular camptothecinconjugate PNU-166148. Irinotecan can be administered, e.g. in the formas it is marketed, e.g. under the trademark Camptosar™. Topotecan ismarketed under the trade name Hycamptin™.

The term “topoisomerase II inhibitor” as used herein includes, but isnot limited to the anthracyclines such as doxorubicin (includingliposomal formulation, such as Caelyx™) daunorubicin, epirubicin,idarubicin and nemorubicin, the anthraquinones mitoxantrone andlosoxantrone, and the podophillotoxines etoposide and teniposide.Etoposide is marketed under the trade name Etopophos™. Teniposide ismarketed under the trade name VM 26-Bristol Doxorubicin is marketedunder the trade name Acriblastin™ or Adriamycin™. Epirubicin is marketedunder the trade name Farmorubicin™. Idarubicin is marketed. under thetrade name Zavedos™. Mitoxantrone is marketed under the trade nameNovantron.

The term “microtubule active agent” relates to microtubule stabilizing,microtubule destabilizing compounds and microtublin polymerizationinhibitors including, but not limited to taxanes, such as paclitaxel anddocetaxel; vinca alkaloids, such as vinblastine or vinblastine sulfate,vincristine or vincristine sulfate, and vinorelbine; discodermolides;cochicine and epothilones and derivatives thereof. Paclitaxel ismarketed under the trade name Taxol™. Docetaxel is marketed under thetrade name Taxotere™. Vinblastine sulfate is marketed under the tradename Vinblastin R.P™. Vincristine sulfate is marketed under the tradename Farmistin™.

The term “alkylating agent” as used herein includes, but is not limitedto, cyclophosphamide, ifosfamide, melphalan or nitrosourea (BCNU orGliadel). Cyclophosphamide is marketed under the trade name Cyclostin™.Ifosfamide is marketed under the trade name Holoxan™.

The term “histone deacetylase inhibitors” or “HDAC inhibitors” relatesto compounds which inhibit the histone deacetylase and which possessantiproliferative activity. This includes, but is not limited to,suberoylanilide hydroxamic acid (SAHA).

The term “antineoplastic antimetabolite” includes, but is not limitedto, 5-fluorouracil or 5-FU, capecitabine, gemcitabine, DNA demethylatingcompounds, such as 5-azacytidine and decitabine, methotrexate andedatrexate, and folic acid antagonists such as pemetrexed. Capecitabineis marketed under the trade name Xeloda™. Gemcitabine is marketed underthe trade name Gemzar™.

The term “platin compound” as used herein includes, but is not limitedto, carboplatin, cis-platin, cisplatinum and oxaliplatin. Carboplatincan be administered, e.g., in the form as it is marketed, e.g. under thetrademark Carboplat™. Oxaliplatin can be administered, e.g., in the formas it is marketed, e.g. under the trademark Eloxatin™.

The term “compounds targeting/decreasing a protein or lipid kinaseactivity; or a protein or lipid phosphatase activity; or furtheranti-angiogenic compounds” as used herein includes, but is not limitedto, protein tyrosine kinase and/or serine and/or threonine kinaseinhibitors or lipid kinase inhibitors, such as a) compounds targeting,decreasing or inhibiting the activity of the platelet-derived growthfactor-receptors (PDGFR), such as compounds which target, decrease orinhibit the activity of PDGFR, especially compounds which inhibit thePDGF receptor, such as an N-phenyl-2-pyrimidine-amine derivative, suchas imatinib, SU101, SU6668 and GFB-111; b) compounds targeting,decreasing or inhibiting the activity of the fibroblast growthfactor-receptors (FGFR); c) compounds targeting, decreasing orinhibiting the activity of the insulin-like growth factor receptor I(IGF-IR), such as compounds which target, decrease or inhibit theactivity of IGF-IR, especially compounds which inhibit the kinaseactivity of IGF-I receptor, or antibodies that target the extracellulardomain of IGF-I receptor or its growth factors; d) compounds targeting,decreasing or inhibiting the activity of the Trk receptor tyrosinekinase family, or ephrin B4 inhibitors; e) compounds targeting,decreasing or inhibiting the activity of the AxI receptor tyrosinekinase family; f) compounds targeting, decreasing or inhibiting theactivity of the Ret receptor tyrosine kinase; g) compounds targeting,decreasing or inhibiting the activity of the Kit/SCFR receptor tyrosinekinase, such as imatinib; h) compounds targeting, decreasing orinhibiting the activity of the C-kit receptor tyrosine kinases, whichare part of the PDGFR family, such as compounds which target, decreaseor inhibit the activity of the c-Kit receptor tyrosine kinase family,especially compounds which inhibit the c-Kit receptor, such as imatinib;i) compounds targeting, decreasing or inhibiting the activity of membersof the c-Abl family, their gene-fusion products (e.g. BCR-Abl kinase)and mutants, such as compounds which target decrease or inhibit theactivity of c-Abl family members and their gene fusion products, such asan N-phenyl-2-pyrimidine-amine derivative, such as imatinib or nilotinib(AMN107); PD180970; AG957; NSC 680410; PD173955 from ParkeDavis; ordasatinib (BMS-354825); j) compounds targeting, decreasing or inhibitingthe activity of members of the protein kinase C (PKC) and Raf family ofserine/threonine kinases, members of the MEK, SRC, JAK/pan-JAK, FAK,PDK1, PKB/Akt, Ras/MAPK, PI3K, SYK, TYK2, BTK and TEC family, and/ormembers of the cyclin-dependent kinase family (CDK) includingstaurosporine derivatives, such as midostaurin; examples of furthercompounds include UCN-01, safingol, BAY 43-9006, Bryostatin 1,Perifosine; llmofosine; RO 318220 and RO 320432; GO 6976; lsis 3521;LY333531/LY379196; isochinoline compounds; FTIs; PD184352 or QAN697 (aP13K inhibitor) or AT7519 (CDK inhibitor); k) compounds targeting,decreasing or inhibiting the activity of protein-tyrosine kinaseinhibitors, such as compounds which target, decrease or inhibit theactivity of protein-tyrosine kinase inhibitors include imatinib mesylate(Gleevec™) or tyrphostin such as Tyrphostin A23/RG-50810; AG 99;Tyrphostin AG 213; Tyrphostin AG 1748; Tyrphostin AG 490; TyrphostinB44; Tyrphostin B44 (+) enantiomer; Tyrphostin AG 555; AG 494;Tyrphostin AG 556, AG957 and adaphostin(4-{[(2,5-dihydroxyphenyl)methyl]amino}-benzoic acid adamantyl ester;NSC 680410, adaphostin); 1) compounds targeting, decreasing orinhibiting the activity of the epidermal growth factor family ofreceptor tyrosine kinases (EGFR₁ ErbB2, ErbB3, ErbB4 as homo- orheterodimers) and their mutants, such as compounds which target,decrease or inhibit the activity of the epidermal growth factor receptorfamily are especially compounds, proteins or antibodies which inhibitmembers of the EGF receptor tyrosine kinase family, such as EGFreceptor, ErbB2, ErbB3 and ErbB4 or bind to EGF or EGF related ligands,CP 358774, ZD 1839, ZM 105180; trastuzumab (Herceptin™), cetuximab(Erbitux™), Iressa, Tarceva, OSI-774, CI-1033, EKB-569, GW-2016, E1.1,E2.4, E2.5, E6.2, E6.4, E2.11, E6.3 or E7.6.3, and7H-pyrrolo-[2,3-d]pyrimidine derivatives; m) compounds targeting,decreasing or inhibiting the activity of the c-Met receptor, such ascompounds which target, decrease or inhibit the activity of c-Met,especially compounds which inhibit the kinase activity of c-Metreceptor, or antibodies that target the extracellular domain of c-Met orbind to HGF, n) compounds targeting, decreasing or inhibiting the kinaseactivity of one or more JAK family members (JAK1/JAK2/JAK3/TYK2 and/orpan-JAK), including but not limited to PRT-062070, SB-1578, baricitinib,pacritinib, momelotinib, VX-509, AZD-1480, TG-101348, tofacitinib, andruxolitinib; o) compounds targeting, decreasing or inhibiting the kinaseactivity of PI3 kinase (PI3K) including but not limited to ATU-027,SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474, buparlisib,pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147, XL-765, andidelalisib; and; and q) compounds targeting, decreasing or inhibitingthe signaling effects of hedgehog protein (Hh) or smoothened receptor(SMO) pathways, including but not limited to cyclopamine, vismodegib,itraconazole, erismodegib, and IPI-926 (saridegib).

The term “PI3K inhibitor” as used herein includes, but is not limited tocompounds having inhibitory activity against one or more enzymes in thephosphatidylinositol-3-kinase family, including, but not limited toPI3Kα, PI3Kγ, PI3Kδ, PI3Kβ, PI3K-C2α, PI3K-C2β, PI3K-C2γ, Vps34, p110-α,p110-β, p110-γ, p110-δ, p85-α, p85-β, p55-γ, p150, p101, and p87.Examples of PI3K inhibitors useful in this invention include but are notlimited to ATU-027, SF-1126, DS-7423, PBI-05204, GSK-2126458, ZSTK-474,buparlisib, pictrelisib, PF-4691502, BYL-719, dactolisib, XL-147,XL-765, and idelalisib.

The term “Bcl-2 inhibitor” as used herein includes, but is not limitedto compounds having inhibitory activity against B-cell lymphoma 2protein (Bcl-2), including but not limited to ABT-199, ABT-731, ABT-737,apogossypol, Ascenta's pan-Bcl-2 inhibitors, curcumin (and analogsthereof), dual Bcl-2/Bcl-xL inhibitors (InfinityPharmaceuticals/Novartis Pharmaceuticals), Genasense (G3139), HA14-1(and analogs thereof; see WO2008118802), navitoclax (and analogsthereof, see U.S. Pat. No. 7,390,799), NH-1 (Shenayng PharmaceuticalUniversity), obatoclax (and analogs thereof, see WO2004106328), S-001(Gloria Pharmaceuticals), TW series compounds (Univ. of Michigan), andvenetoclax. In some embodiments the Bcl-2 inhibitor is a small moleculetherapeutic. In some embodiments the Bcl-2 inhibitor is apeptidomimetic.

The term “BTK inhibitor” as used herein includes, but is not limited tocompounds having inhibitory activity against Bruton's Tyrosine Kinase(BTK), including, but not limited to AVL-292 and ibrutinib.

The term “SYK inhibitor” as used herein includes, but is not limited tocompounds having inhibitory activity against spleen tyrosine kinase(SYK), including but not limited to PRT-062070, R-343, R-333, Excellair,PRT-062607, and fostamatinib.

Further examples of BTK inhibitory compounds, and conditions treatableby such compounds in combination with compounds of this invention can befound in WO2008039218 and WO2011090760, the entirety of which areincorporated herein by reference.

Further examples of SYK inhibitory compounds, and conditions treatableby such compounds in combination with compounds of this invention can befound in WO2003063794, WO2005007623, and WO2006078846, the entirety ofwhich are incorporated herein by reference.

Further examples of PI3K inhibitory compounds, and conditions treatableby such compounds in combination with compounds of this invention can befound in WO2004019973, WO2004089925, WO2007016176, U.S. Pat. No.8,138,347, WO2002088112, WO2007084786, WO2007129161, WO2006122806,WO2005113554, and WO2007044729 the entirety of which are incorporatedherein by reference.

Further examples of JAK inhibitory compounds, and conditions treatableby such compounds in combination with compounds of this invention can befound in WO2009114512, WO2008109943, WO2007053452, WO2000142246, andWO2007070514, the entirety of which are incorporated herein byreference.

Further anti-angiogenic compounds include compounds having anothermechanism for their activity, e.g. unrelated to protein or lipid kinaseinhibition e.g. thalidomide (Thalomid™) and TNP-470.

Examples of proteasome inhibitors useful for use in combination withcompounds of the invention include, but are not limited to bortezomib,disulfiram, epigallocatechin-3-gallate (EGCG), salinosporamide A,carfilzomib, ONX-0912, CEP-18770, and MLN9708.

Compounds which target, decrease or inhibit the activity of a protein orlipid phosphatase are e.g. inhibitors of phosphatase 1, phosphatase 2A,or CDC25, such as okadaic acid or a derivative thereof.

Compounds which induce cell differentiation processes include, but arenot limited to, retinoic acid, α- γ- or δ-tocopherol or α- γ- orδ-tocotrienol.

The term cyclooxygenase inhibitor as used herein includes, but is notlimited to, Cox-2 inhibitors, 5-alkyl substituted2-arylaminophenylacetic acid and derivatives, such as celecoxib(Celebrex™), rofecoxib (Vioxx™), etoricoxib, valdecoxib or a 5-alkyl-2-arylaminophenylacetic acid, such as5-methyl-2-(2′-chloro-6′-fluoroanilino)phenyl acetic acid, lumiracoxib.

The term “bisphosphonates” as used herein includes, but is not limitedto, etridonic, clodronic, tiludronic, pamidronic, alendronic,ibandronic, risedronic and zoledronic acid. Etridonic acid is marketedunder the trade name Didronel™. Clodronic acid is marketed under thetrade name Bonefos™. Tiludronic acid is marketed under the trade nameSkelid™ Pamidronic acid is marketed under the trade name Aredia™.Alendronic acid is marketed under the trade name Fosamax™. Ibandronicacid is marketed under the trade name Bondranat™ Risedronic acid ismarketed under the trade name Actonel™. Zoledronic acid is marketedunder the trade name Zometa™. The term “mTOR inhibitors” relates tocompounds which inhibit the mammalian target of rapamycin (mTOR) andwhich possess antiproliferative activity such as sirolimus (Rapamune®),everolimus (Certican™), CCI-779 and ABT578.

The term “heparanase inhibitor” as used herein refers to compounds whichtarget, decrease or inhibit heparin sulfate degradation. The termincludes, but is not limited to, PI-88. The term “biological responsemodifier” as used herein refers to a lymphokine or interferons.

The term “inhibitor of Ras oncogenic isoforms”, such as H-Ras, K-Ras, orN-Ras, as used herein refers to compounds which target, decrease orinhibit the oncogenic activity of Ras; for example, a “farnesyltransferase inhibitor” such as L-744832, DK8G557 or R115777(Zarnestra™). The term “telomerase inhibitor” as used herein refers tocompounds which target, decrease or inhibit the activity of telomerase.Compounds which target, decrease or inhibit the activity of telomeraseare especially compounds which inhibit the telomerase receptor, such astelomestatin.

The term “methionine aminopeptidase inhibitor” as used herein refers tocompounds which target, decrease or inhibit the activity of methionineaminopeptidase. Compounds which target, decrease or inhibit the activityof methionine aminopeptidase include, but are not limited to, bengamideor a derivative thereof.

The term “proteasome inhibitor” as used herein refers to compounds whichtarget, decrease or inhibit the activity of the proteasome. Compoundswhich target, decrease or inhibit the activity of the proteasomeinclude, but are not limited to, Bortezomib (Velcade™) and MLN 341.

The term “matrix metalloproteinase inhibitor” or (“MMP” inhibitor) asused herein includes, but is not limited to, collagen peptidomimetic andnonpeptidomimetic inhibitors, tetracycline derivatives, e.g. hydroxamatepeptidomimetic inhibitor batimastat and its orally bioavailable analoguemarimastat (BB-2516), prinomastat (AG3340), metastat (NSC 683551)BMS-279251, BAY 12-9566, TAA211, MMI270B or AAJ996.

The term “compounds used in the treatment of hematologic malignancies”as used herein includes, but is not limited to, FMS-like tyrosine kinaseinhibitors, which are compounds targeting, decreasing or inhibiting theactivity of FMS-like tyrosine kinase receptors (Flt-3R); interferon,1-β-D-arabinofuransylcytosine (ara-c) and bisulfan; and ALK inhibitors,which are compounds which target, decrease or inhibit anaplasticlymphoma kinase.

Compounds which target, decrease or inhibit the activity of FMS-liketyrosine kinase receptors (Flt-3R) are especially compounds, proteins orantibodies which inhibit members of the Flt-3R receptor kinase family,such as PKC412, midostaurin, a staurosporine derivative, SU11248 andMLN518.

The term “HSP90 inhibitors” as used herein includes, but is not limitedto, compounds targeting, decreasing or inhibiting the intrinsic ATPaseactivity of HSP90; degrading, targeting, decreasing or inhibiting theHSP90 client proteins via the ubiquitin proteosome pathway. Compoundstargeting, decreasing or inhibiting the intrinsic ATPase activity ofHSP90 are especially compounds, proteins or antibodies which inhibit theATPase activity of HSP90, such as 17-allylamino,17-demethoxygeldanamycin(17AAG), a geldanamycin derivative; other geldanamycin relatedcompounds; radicicol and HDAC inhibitors.

The term “antiproliferative antibodies” as used herein includes, but isnot limited to, trastuzumab (Herceptin™), Trastuzumab-DM1, erbitux,bevacizumab (Avastin™), rituximab (Rituxan®), PRO64553 (anti-CD40) and2C4 Antibody. By antibodies is meant intact monoclonal antibodies,polyclonal antibodies, multispecific antibodies formed from at least 2intact antibodies, and antibodies fragments so long as they exhibit thedesired biological activity.

For the treatment of acute myeloid leukemia (AML), compounds of thecurrent invention can be used in combination with standard leukemiatherapies, especially in combination with therapies used for thetreatment of AML. In particular, compounds of the current invention canbe administered in combination with, for example, farnesyl transferaseinhibitors and/or other drugs useful for the treatment of AML, such asDaunorubicin, Adriamycin, Ara-C, VP-16, Teniposide, Mitoxantrone,Idarubicin, Carboplatinum and PKC412.

Other anti-leukemic compounds include, for example, Ara-C, a pyrimidineanalog, which is the 2′-alpha-hydroxy ribose (arabinoside) derivative ofdeoxycytidine. Also included is the purine analog of hypoxanthine,6-mercaptopurine (6-MP) and fludarabine phosphate. Compounds whichtarget, decrease or inhibit activity of histone deacetylase (HDAC)inhibitors such as sodium butyrate and suberoylanilide hydroxamic acid(SAHA) inhibit the activity of the enzymes known as histonedeacetylases. Specific HDAC inhibitors include MS275, SAHA, FK228(formerly FR901228), Trichostatin A and compounds disclosed in U.S. Pat.No. 6,552,065 including, but not limited to,N-hydroxy-3-[4-[[[2-(2-methyl-1H-indol-3-yl)-ethyl]-amino]methyl]phenyl]-2E-2-propenamide,or a pharmaceutically acceptable salt thereof andN-hydroxy-3-[4-[(2-hydroxyethyl){2-(1H-indol-3-yl)ethyl]-amino]methyl]phenyl]-2E-2-propenamide,or a pharmaceutically acceptable salt thereof, especially the lactatesalt. Somatostatin receptor antagonists as used herein refer tocompounds which target, treat or inhibit the somatostatin receptor suchas octreotide, and SOM230. Tumor cell damaging approaches refer toapproaches such as ionizing radiation. The term “ionizing radiation”referred to above and hereinafter means ionizing radiation that occursas either electromagnetic rays (such as X-rays and gamma rays) orparticles (such as alpha and beta particles). Ionizing radiation isprovided in, but not limited to, radiation therapy and is known in theart. See Hellman, Principles of Radiation Therapy, Cancer, in Principlesand Practice of Oncology, Devita et al., Eds., 4^(th) Edition, Vol. 1,pp. 248-275 (1993).

Also included are EDG binders and ribonucleotide reductase inhibitors.The term “EDG binders” as used herein refers to a class ofimmunosuppressants that modulates lymphocyte recirculation, such asFTY720. The term “ribonucleotide reductase inhibitors” refers topyrimidine or purine nucleoside analogs including, but not limited to,fludarabine and/or cytosine arabinoside (ara-C), 6-thioguanine,5-fluorouracil, cladribine, 6-mercaptopurine (especially in combinationwith ara-C against ALL) and/or pentostatin. Ribonucleotide reductaseinhibitors are especially hydroxyurea or2-hydroxy-1H-isoindole-1,3-dione derivatives.

Also included are in particular those compounds, proteins or monoclonalantibodies of VEGF such as1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine or a pharmaceuticallyacceptable salt thereof,1-(4-chloroanilino)-4-(4-pyridylmethyl)phthalazine succinate;Angiostatin™; Endostatin™; anthranilic acid amides; ZD4190; Zd₆474;SU5416; SU6668; bevacizumab; or anti-VEGF antibodies or anti-VEGFreceptor antibodies, such as rhuMAb and RHUFab, VEGF aptamer such asMacugon; FLT-4 inhibitors, FLT-3 inhibitors, VEGFR-2 IgGI antibody,Angiozyme (RPI 4610) and Bevacizumab (Avastin™)

Photodynamic therapy as used herein refers to therapy which uses certainchemicals known as photosensitizing compounds to treat or preventcancers. Examples of photodynamic therapy include treatment withcompounds, such as Visudyne™ and porfimer sodium.

Angiostatic steroids as used herein refers to compounds which block orinhibit angiogenesis, such as, e.g., anecortave, triamcinolone,hydrocortisone, 11-α-epihydrocotisol, cortexolone,17α-hydroxyprogesterone, corticosterone, desoxycorticosterone,testosterone, estrone and dexamethasone.

Implants containing corticosteroids refers to compounds, such asfluocinolone and dexamethasone.

Other chemotherapeutic compounds include, but are not limited to, plantalkaloids, hormonal compounds and antagonists; biological responsemodifiers, preferably lymphokines or interferons; antisenseoligonucleotides or oligonucleotide derivatives; shRNA or siRNA; ormiscellaneous compounds or compounds with other or unknown mechanism ofaction.

The structure of the active compounds identified by code numbers,generic or trade names may be taken from the actual edition of thestandard compendium “The Merck Index” or from databases, e.g. PatentsInternational (e.g. IMS World Publications).

A compound of the current invention may also be used in combination withknown therapeutic processes, for example, the administration of hormonesor radiation. In certain embodiments, a provided compound is used as aradiosensitizer, especially for the treatment of tumors which exhibitpoor sensitivity to radiotherapy.

A compound of the current invention can be administered alone or incombination with one or more other therapeutic compounds, possiblecombination therapy taking the form of fixed combinations or theadministration of a compound of the invention and one or more othertherapeutic compounds being staggered or given independently of oneanother, or the combined administration of fixed combinations and one ormore other therapeutic compounds. A compound of the current inventioncan besides or in addition be administered especially for tumor therapyin combination with chemotherapy, radiotherapy, immunotherapy,phototherapy, surgical intervention, or a combination of these.Long-term therapy is equally possible as is adjuvant therapy in thecontext of other treatment strategies, as described above. Otherpossible treatments are therapy to maintain the patient's status aftertumor regression, or even chemopreventive therapy, for example inpatients at risk.

Those additional agents may be administered separately from an inventivecompound-containing composition, as part of a multiple dosage regimen.Alternatively, those agents may be part of a single dosage form, mixedtogether with a compound of this invention in a single composition. Ifadministered as part of a multiple dosage regime, the two active agentsmay be submitted simultaneously, sequentially or within a period of timefrom one another normally within five hours from one another.

As used herein, the term “combination,” “combined,” and related termsrefers to the simultaneous or sequential administration of therapeuticagents in accordance with this invention. For example, a compound of thepresent invention may be administered with another therapeutic agentsimultaneously or sequentially in separate unit dosage forms or togetherin a single unit dosage form. Accordingly, the present inventionprovides a single unit dosage form comprising a compound of the currentinvention, an additional therapeutic agent, and a pharmaceuticallyacceptable carrier, adjuvant, or vehicle.

The amount of both an inventive compound and additional therapeuticagent (in those compositions which comprise an additional therapeuticagent as described above) that may be combined with the carriermaterials to produce a single dosage form will vary depending upon thehost treated and the particular mode of administration. Preferably,compositions of this invention should be formulated so that a dosage ofbetween 0.01-100 mg/kg body weight/day of an inventive compound can beadministered.

In those compositions which comprise an additional therapeutic agent,that additional therapeutic agent and the compound of this invention mayact synergistically. Therefore, the amount of additional therapeuticagent in such compositions will be less than that required in amonotherapy utilizing only that therapeutic agent. In such compositionsa dosage of between 0.01-1,000 μg/kg body weight/day of the additionaltherapeutic agent can be administered.

The amount of additional therapeutic agent present in the compositionsof this invention will be no more than the amount that would normally beadministered in a composition comprising that therapeutic agent as theonly active agent. Preferably the amount of additional therapeutic agentin the presently disclosed compositions will range from about 50% to100% of the amount normally present in a composition comprising thatagent as the only therapeutically active agent.

The compounds of this invention, or pharmaceutical compositions thereof,may also be incorporated into compositions for coating an implantablemedical device, such as prostheses, artificial valves, vascular grafts,stents and catheters. Vascular stents, for example, have been used toovercome restenosis (re-narrowing of the vessel wall after injury).However, patients using stents or other implantable devices risk clotformation or platelet activation. These unwanted effects may beprevented or mitigated by pre-coating the device with a pharmaceuticallyacceptable composition comprising a kinase inhibitor. Implantabledevices coated with a compound of this invention are another embodimentof the present invention.

EXEMPLIFICATION

General Synthetic Methods

The following examples are intended to illustrate the invention and arenot to be construed as being limitations thereon. Unless otherwisestated, one or more tautomeric forms of compounds of the examplesdescribed hereinafter may be prepared in situ and/or isolated. Alltautomeric forms of compounds of the examples described hereafter shouldbe considered to be disclosed. Temperatures are given in degreescentigrade. If not mentioned otherwise, all evaporations are performedunder reduced pressure, preferably between about 15 mm Hg and 100 mm Hg(=20-133 mbar). The structure of final products, intermediates andstarting materials is confirmed by standard analytical methods, e.g.,microanalysis and spectroscopic characteristics, e.g., MS, IR, NMR.Abbreviations used are those conventional in the art.

All starting materials, building blocks, reagents, acids, bases,dehydrating agents, solvents, and catalysts utilized to synthesis thecompounds of the present invention are either commercially available orcan be produced by organic synthesis methods known to one of ordinaryskill in the art (Houben-Weyl 4th Ed. 1952, Methods of OrganicSynthesis, Thieme, Volume 21). Further, the compounds of the presentinvention can be produced by organic synthesis methods known to one ofordinary skill in the art as shown in the following examples.

As depicted in the Examples below, in certain exemplary embodiments,compounds are prepared according to the following general procedures. Itwill be appreciated that, although the general methods depict thesynthesis of certain compounds of the present invention, the followinggeneral methods, and other methods known to one of ordinary skill in theart, can be applied to all compounds and subclasses and species of eachof these compounds, as described herein.

ABBREVIATIONS

equiv or eq: molar equivalentso/n: overnightrt: room temperatureUV: ultra violetHPLC: high pressure liquid chromatographyRt: retention timeLCMS or LC-MS: liquid chromatography-mass spectrometryNMR: nuclear magnetic resonanceCC: column chromatographyTLC: thin layer chromatographysat: saturatedaq: aqueousAc: acetylDCM: dichloromethaneDCE: dichloroethaneDEA: diethylamineDMF: dimethylformamideDMSO: dimethylsulfoxideACN or MeCN: acetonitrileDIPEA: diisopropylethylamineEA or EtOAc: ethyl acetateBINAP: (±)-2,2′-Bis(diphenylphosphino)-1,1′-binaphthaleneTEA: triethylamineTHF: tetrahydrofuranTBS: tert-butyldimethylsilylKHMDS: potassium hexamethyl disilylazideTf: trifluoromethanesulfonateMs: methanesulfonyl

NBS: N-bromosuccinimide

PE: petroleum etherTFA: trifluoroacetic acidMMPP: magnesium monoperoxyphthalateHATU: 1-[Bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium3-oxid

Hexafluorophosphate NCS: N-chlorosuccinimide

Cy: cyclohexylTol: tolueneDMP: Dess-Martin periodinaneIBX: 2-iodoxybenzoic acidPMB: p-methoxybenzylSEM: [2-(Trim ethyl silyl)ethoxy]methylXPhos or X-Phos: 2-Dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl

General information: All evaporations were carried out in vacuo with arotary evaporator. Analytical samples were dried in vacuo (1-5 mmHg) atrt. Thin layer chromatography (TLC) was performed on silica gel plates,spots were visualized by UV light (214 and 254 nm). Purification bycolumn and flash chromatography was carried out using silica gel(200-300 mesh). Solvent systems are reported as mixtures by volume. All¹H NMR spectra were recorded on a Bruker 400 (400 MHz) spectrometer. ¹Hchemical shifts are reported in δ values in parts per million (ppm) withthe deuterated solvent as the internal standard. Data are reported asfollows: chemical shift, multiplicity (s=singlet, d=doublet, t=triplet,q=quartet, br=broad, m=multiplet), coupling constant (Hz), integration(i.e. number of protons). LCMS spectra were obtained on an Agilent 1200series 6110 or 6120 mass spectrometer with electrospray ionization andexcept as otherwise indicated, the general LCMS conditions were asfollows: Waters X Bridge C18 column (50 mm*4.6 mm*3.5 μm), Flow Rate:2.0 mL/min, the column temperature: 40° C.

General Procedure a (Wolff-Kishner Reduction):

A mixture of 2,6-diaryl piperidin-4-one (concentration 0.1-1 M), KOH (20eq.) and N₂H₄.H₂O (40 eq.) in diethylene glycol was stirred for about 2hours at 80° C. and then at approx. 150-200° C. until the reactioncompleted. After cooled down to room temperature, the reaction mixturewas diluted with water and extracted with DCM. The organic layer waswashed with water and brine, dried over Na₂SO₄, filtered andconcentrated in vacuum. The residue was purified by columnchromatography to give 2,6-diaryl piperidine.

General Procedure B (N-Alkylation of 2,6-Diaryl Piperidine):

To a solution of 2,6-diaryl piperidine (concentration 0.1-1 M) in DMF orACN was added corresponding halide or mesylate (2 eq.) and K₂CO₃ (2 eq.)under Ar atmosphere. The mixture was stirred at 80° C. overnight, thenit was diluted with H₂O and extracted with DCM. The combined organiclayers were washed with water, dried over Na₂SO₄, filtered andconcentrated in vacuo to give desired N-alkylated target.

General Procedure C (Reaction of Alcohols with MethanesulfonylChloride):

To a solution of alcohol (concentration 0.1-1 M) and Et3N (approx. 2.5eq.) in DCM was added MSCI (1.2-1.4 eq.) drop wise at −70° C., and thereaction mixture was stirred at room temperature for 30 mins, then theresulting mixture was quenched with NaHCO₃aq. and extracted with DCM.The combined organic layers were washed with water and brine, dried overNa₂SO₄ and filtered. The filtrate was concentrated in vacuum to give thecorresponding mesylate.

General Procedure D (Reaction of Mesylates or Halides with 2,6-DiarylPiperidine):

A mixture of 2,6-diaryl piperidine (concentration 0.1-1 M),corresponding mesylate or halide (approx. 2-3 eq.), KI (0.2-0.3 eq.),DIPEA (2-3 eq.) in DMF or ACN was stirred overnight at 60-80° C. andfiltered. The filtrate was purified by prep-HPLC to get the alkylated2,6-diaryl piperidine.

General Procedure E (Reaction of Aryl Aldehyde with Acetone to Give4-(heteroaryl or aryl)but-3-en-2-one):

A mixture of corresponding aryl aldehyde (concentration 0.1-1 M),acetone (20 eq.) and K₂CO₃ (1.5-2 eq.) in toluene/EtOH/H₂O (5:2:1) wasstirred at 80° C. for approx. 13 hours and cooled down to roomtemperature. After diluted with EA, The reaction mixture was filteredthrough basic silica gel column and washed with DCM/MeOH (100/1). Thefiltrate was concentrated in vacuum to give 4-(heteroaryl oraryl)but-3-en-2-one which was used in the next step without furtherpurification.

General Procedure F (Reaction of Aryl Aldehyde with Acetone to Give4-(heteroaryl or aryl)but-3-en-2-one):

To a mixture of aryl aldehyde (concentration 0.1-1 M) in acetone wereadded a solution of NaOH (approx. 8 M, 1.5 eq.) in H₂O at 0° C. Themixture was stirred at 0° C. for 1 hour. Then it was warmed to roomtemperature and stirred another 2 hours. The solution was adjusted pH to8 with 35% HCl aq., dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by column chromatographyto give 4-(heteroaryl or aryl)but-3-en-2-one.

General Procedure G (Buchwald Coupling of Aryl Bromide with AlkylAmine):

A mixture of aryl bromide (concentration 0.1-1 M), alkyl amine (2 eq,0.2-2 M), Pd(OAc)₂ (0.1-0.15 eq), BINAP (0.2-0.3 eq), and Cs2CO3 (2-4eq) in toluene was stirred at 75-120° C. overnight. After completed, thereaction mixture was concentrated in vacuum and purified by columnchromatography to afford the desired product.

General Procedure H (Suzuki Coupling of Aryl Bromide with Aryl BoronicAcid):

aryl bromide (concentration 0.1-1 M), aryl boronic acid (1.1-1.5 eq),PdC12(dppf) (0.05-0.08 eq), and Na2CO3 aq. (1 M, 2.5 eq) in 1,4-dioxanewas stirred at 80-100° C. for 10 mins under microwave irradiation. Afterthe reaction was completed, the mixture was diluted with water and theaqueous layer was extracted with DCM 3 times. The combined organiclayers were washed with brine, dried over Na₂SO₄ and filtered. Thefiltrate was concentrated in vacuum and the residue was purified bysilica gel column.

General Procedure I (Reductive Amination of Secondary Amine to TertiaryAmine):

To a mixture of secondary amine (concentration 0.1-1 M), correspondingaldehyde or ketone (1-2 eq) and NaBH(OAc)₃ (3-6 eq) in DCM was addedseveral drops of acetic acid, and then the mixture was stirred at roomtemperature for 2-18 h. The mixture was neutralized with saturatedNaHCO₃ aqueous solution to pH=8-9 and extracted with DCM. The organiclayers were washed with brine, dried over Na₂SO₄, filtered andconcentrated in vacuum to give the desired tertiary amine.

General Procedure J (Boc Cleavage of N-Boc Protected Amines):

To a solution of N-Boc protected amine (concentration 0.1-1 M) in DCMwas added TFA (1/15 volume of DCM) at room temperature. The reactionmixture was stirred for 2 h, then concentrated and saturated NaHCO₃aqueous solution was added and the mixture was extracted with DCM. Theorganic extracts were dried over Na₂SO₄, filtered and concentrated togive the free amine as the desired product.

General Procedure K (Halogenation of imidazo[1,2-a]pyridine to Give3-Halogenated imidazo[1,2-a]pyridine):

A mixture of imidazo[1,2-a]pyridine derivatives (concentration 0.1-1 M)and NBS or NCS (0.8-0.9 eq) in DCM (10 mL) was stirred at roomtemperature for 1 hour. After the reaction was completed, the suspensionwas diluted with water and DCM, the separated organic layer wasconcentrated by vacuum and the residue was purified by prep-HPLC to givethe desired product.

Example 1: Synthesis of I-1 and I-3

The synthesis of X4-014-H:

Following general procedure E, X4-014-H (1.9 g, 41%) was obtained asyellow foam, which was used in the next step without furtherpurification. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:69.13%; Rt=1.38 min; MS Calcd.: 284.4; MS Found: 285.4[M+H]⁺.

The synthesis of I-3:

To a solution of X4-014-H (1.4 g, 4.9 mmol) in MeOH (20 mL) was addedL-proline (227 mg, 1.97 mmol), 3-methylpicolinaldehyde (656 mg, 5.4mmol) and aq. MeNH₂ (1.5 g, 40% wt, 19.72 mmol) sequentially. Thereaction mixture was stirred overnight at room temperature andconcentrated in vacuum. The residue was purified by columnchromatography to give cis/trans mixture of I-3 (mg, 50%) as a yellowfoam, which was used in the next step without further purification.Cis/trans mixture of I-3 (270 mg, 0.65 mmol) was purified by prep-TLC togive I-3 (30 mg, 11%) as a white solid. LCMS (Agilent LCMS 1200-6120,Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in1.6 min, then under this condition for 1.4 min, finally changed to 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this conditionfor 0.7 min). I-3: Purity: 92.86%. Rt=1.49 min (trans), 1.52 min (cis);MS Calcd.: 418.3; MS Found: 419.4 [M+H]⁺. HPLC (Agilent HPLC 1200,Column: Waters X-Bridge C18 (150 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in10 min, then under this condition for 5 min, finally changed to 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this conditionfor 5 min.); Purity: 97.88%. Rt=4.90 min. ¹H NMR (400 MHz, CDCl₃) δ:8.58-8.56 (m, 1H), 7.62 (s, 1H), 7.48 (dd, J=1.2 Hz, J=7.6 Hz, 1H), 7.36(d, J=8.8 Hz 1H), 7.19 (dd, J=7.2 Hz, J=8.8 Hz, 1H), 7.13 (dd, J=4.8 Hz,J=7.6 Hz, 1H), 6.34 (d, J=6.4 Hz, 1H), 4.03 (dd, J=3.2 Hz, J=12.0 Hz,1H), 3.93 (dd, J=7.2 Hz, J=8.8 Hz, 1H), 3.49 (s, 1H), 3.33-3.18 (m, 6H),2.78-2.68 (m, 5H), 2.50 (s, 3H), 2.42 (s, 3H), 1.94 (s, 3H).

Synthesis of I-1:

Following general procedure A, a mixture of cis/trans mixture of I-3(770 mg, 1.84 mmol), KOH (2.1 g, 36.8 mmol) and N₂H₄.H₂O (4.6 g, 80% wt,73.6 mmol) in diethylene glycol was stirred for 2 hours at 80° C. andthen 5 hours at 150° C. After cooled down to room temperature, thereaction mixture was diluted with water and extracted with DCM. Theorganic layer was washed with water and brine, dried over Na₂SO₄,filtered and concentrated in vacuum. The residue was purified by columnchromatography to give 276 mg of crude I-1, which was purified byprep-HPLC to give 80 mg of I-1 as a white solid. LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 1.6 min, then under this condition for 1.4 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 0.7 min.); Purity: 96.44%. Rt=1.66 min; MS Calcd.:404.3; MS Found: 405.4 [M+H]⁺. HPLC (Agilent HPLC 1200; Column:L-column2 ODS (150 mm*4.6 mm*5.0 μm); Column Temperature: 40° C.; FlowRate: 1.0 mL/min; Mobile Phase: from 95% [water+0.05% TFA] and 5%[CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and 100% [CH₃CN+0.05% TFA] in10 min, then under this condition for 5 min, finally changed to 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1 min and under thiscondition for 5 min.); Purity: 94.20%. Rt=4.44 min. ¹H NMR (400 MHz,CD₃OD) δ: 8.46 (s, 1H), 7.96 (s, 1H), 7.62 (d, J=8.0 Hz, 1H), 7.34-7.25(m, 2H), 7.22-7.19 (m, 1H), 6.51 (d, J=6.8 Hz, 1H), 3.65 (d, J=10.0 Hz,1H), 3.46 (d, J=8.4 Hz, 1H), 3.17 (s, 4H), 2.81 (s, 4H), 2.51 (s, 3H),2.47 (s, 3H), 2.05-1.94 (m, 4H), 1.89 (s, 3H), 1.79-1.68 (m, 2H).

Example 2: Synthesis of I-2

The synthesis of X4-027-A-1:

To a solution of X4-027-A (731 mg, 5.0 mmol) and X4-E (806 mg, 5.0 mmol)in toluene (25 mL), EtOH (10 mL) and H₂O (5 mL) was added K₂CO₃ (1.037g, 7.5 mmol). The mixture was stirred overnight at 70° C. under Aratmosphere. Then it was diluted with H₂O and extracted with DCM threetimes. The combined organic layers were dried over Na₂SO₄, filtered andconcentrated in vacuo. The residue was purified by chromatography ofsilica gel with DCM/MeOH/NH₃—H₂O (40/1/0.68) as eluent to provideproduct X4-027A-1 (952 mg, 66% yield) as yellow solid. LCMS (AgilentLCMS 1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm);Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 1.6 min, then under this condition for 1.4 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 0.7 min.); Purity: 68.23%; Rt=1.55 min; MS Calcd.:298.1; MS Found: 290.1 [M+H]⁺.

The synthesis of X4-027-4:

To a solution of X4-027-A-1 (676 mg, 2.34 mmol) in MeOH (20 mL) wasadded NH₃/H₂O (5 mL). The mixture was stirred at room temperatureovernight. The solvent was evaporated in vacuo and the residue waspurified by prep-TLC to give X4-027-4 (170 mg, 23% yield) as brown-redsolid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min;Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0%[water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:96.20%. Rt=1.41 min; MS Calcd.: 306.1; MS Found: 307.4 [M+H]⁺.

The synthesis of X4-027-5:

Following general procedure A, X4-027-5 (77 mg, 47% yield) was obtainedas brown-red solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min.); Purity: 88.42%. Rt=1.47 min; MS Calcd.: 292.2; MS Found: 293.1[M+H]⁺.

The synthesis of X4-027-6:

Following general procedure B, X4-027-6 (180 mg, >100% yield) wasobtained as white solid, which was used to next step without furtherpurification. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:48.71%. Rt=1.79 min; MS Calcd.: 493.6; MS Found: 494.4 [M+H]⁺.

The synthesis of I-2:

To a solution of crude X4-027-5 (170 mg, 0.25 mmol) in EtOH (3 mL) wasN₂H₄.H₂O (0.6 mL). The mixture was stirred at room temperatureovernight. The solvent was evaporated in vacuo and the residue waspurified by prep-HPLC to give I-2 (12 mg, 13% yield) as white solid.LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; MobilePhase: from 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [(total10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] to 10% [(total 10 mM AcONH₄)H₂O/ACN=9/1 (v/v)] and 90% [(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in1.6 min, then under this condition for 2.4 min, finally changed to 90%[(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [(total 10 mM AcONH₄)H₂O/ACN=1/9 (v/v)] in 0.1 min and under this condition for 0.7 min.);Purity: 98.47%; Rt=1.26 min; MS Calcd.: 363.5; MS Found: 364.3 [M+H]⁺.HPLC (Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm);Column Temperature: 40° C.; Flow Rate: 1.5 mL/min; Mobile Phase: from90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [total 10 mMAcONH₄) H₂O/ACN=1/9 (v/v)] to 15% [total 10 mM AcONH₄) H₂O/ACN=9/1(v/v)] and 85% [total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 5 min, thenunder this condition for 10 min, finally changed to 90% [(total 10 mMAcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [total 10 mM AcONH₄) H₂O/ACN=1/9(v/v)] in 0.1 min and under this condition for 5 min.); Purity: 98.11%.Rt=4.22 min. ¹H NMR (CDCl₃, 400 MHz) δ 8.47 (d, J=4.4 Hz, 1H), 8.08 (d,J=6.8 Hz, 1H), 7.64 (s, 1H), 7.56 (d, J=8.8 Hz, 1H), 7.42 (d, J=7.2 Hz,1H), 7.14-7.10 (m, 1H), 7.06 (dd, J=4.8 Hz, 7.6 Hz, 1H), 6.77-6.73 (m,1H), 4.02-3.99 (m, 1H), 3.90-3.87 (m, 1H), 2.52 (s, 3H), 2.38-2.20 (m,4H), 2.02-1.86 (m, 4H), 1.37-1.29 (m, 2H), 1.13-1.01 (m, 1H), 0.87-0.79(m, 3H).

Example 3: Synthesis of I-4

The synthesis of X4-116-A:

Following general procedure G, X4-116-A (0.92 g, 24% yield) was obtainedas a yellow solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:92.35%; Rt=1.81 min; MS Calcd.: 504.7, MS Found: 505.7 [M+1]

The synthesis of X4-116-B:

Following general procedure A, X4-116-B (140 mg, 16% yield) was obtainedas a light-yellow solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min.); Purity: 93.71%; Rt=1.99 min; MS Calcd.: 490.7, MS Found: 491.7[M+1]

The synthesis of I-4:

Following general procedure J, I-4 (110 mg, 99% yield) was obtained as alight-yellow solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min). Purity: 99.8%; Rt=1.48 min; MS Calcd.: 390.7; MS Found: 391.7[M+H]⁺. HPLC (Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6mm*5.0 μm); Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; MobilePhase: from 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0%[water+0.05% TFA] and 100% [CH₃CN+0.05% TFA] in 10 min, then under thiscondition for 5 min, finally changed to 95% [water+0.05% TFA] and 5%[CH₃CN+0.05% TFA] in 0.1 min and under this condition for 5 min.);Purity: 98.8%; Rt=4.362 min; MS Calcd.: 390.7; MS Found: 391.7 [M+H]⁺.¹H NMR (400 MHz, CDCl₃) δ 8.51 (d, J=3.6 Hz, 1H), 7.63 (s, 1H), 7.43 (d,J=7.2 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 7.12-7.16 (m, 1H), 7.04-7.07 (m,1H), 6.27 (d, J=7.2 Hz, 1H), 3.57 (dd, J=11.2 Hz, J=2.4 Hz, 1H), 3.47(dd, J=10.0 Hz, J=3.6 Hz, 1H), 3.11-3.15 (m, 8H), 2.47 (s, 3H), 2.22 (s,3H), 1.97-2.06 (m, 4H), 1.61-1.75 (m, 2H).

Example 4: Synthesis of I-5, I-6, and I-7

The synthesis of X4-117-1

Following general procedure H, X4-117-1 (63 mg, 32% yield) was obtainedas yellow oil. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)]and 10% [(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] to 10% [(total 10 mMAcONH₄) H₂O/ACN=9/1 (v/v)] and 90% [(total 10 mM AcONH₄) H₂O/ACN=1/9(v/v)] in 1.6 min, then under this condition for 2.4 min, finallychanged to 90% [(total 10 mM AcONH₄) H₂O/ACN=900/100 (v/v)] and 10%[(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 0.1 min and under thiscondition for 0.7 min.); Purity: 54.67%; Rt=1.49 min (trans) & 1.53 min(cis); MS Calcd.: 397.2; MS Found: 398.7[M+H]⁺.

The Preparation of I-5

Following general procedure A, I-5 (13 mg, 21% yield) was obtained asoff-white solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:100.00%; Rt=1.62 min; MS Calcd.: 383.2; MS Found: 384.7[M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 100.00%; Rt=4.70 min;MS Calcd.: 383.2; MS Found: 384.7[M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ1.53-1.65 (m, 2H), 1.81 (s, 3H), 1.83-1.95 (m, 4H), 2.37 (s, 3H),3.37-3.40 (m, 1H), 3.45-3.48 (m, 1H), 6.72 (d, J=6.8 Hz, 1H), 6.96-6.99(m, 1H), 7.15-7.18 (m, 1H), 7.34 (d, J=7.6 Hz, 1H), 7.52 (d, J=6.0 Hz,2H), 7.56 (d, J=9.2 Hz, 1H), 7.71 (s, 1H), 8.42 (d, J=2.4 Hz, 1H), 8.77(d, J=6.0 Hz, 2H).

The Preparation of X4-118-1

Following general procedure H, X4-118-1 (116 mg, 80% yield) was obtainedas yellow solid from X4-101-1 (150 mg, 0.38 mmol). LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 90%[(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [(total 10 mM AcONH₄)H₂O/ACN=1/9 (v/v)] to 10% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and90% [(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 1.6 min, then under thiscondition for 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)H₂O/ACN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)]in 0.1 min and under this condition for 0.7 min.); Purity: 82.37%;Rt=1.63 min (trans) & 1.67 min (cis); MS Calcd.: 397.7; MS Found: 398.7[M+H]⁺.

The Synthesis of I-6

Following general procedure A, I-6 (50 mg, 52% yield) was obtained aswhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:95.04%; Rt=1.64 min; MS Calcd.: 383.7; MS Found: 384.7[M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 100%; Rt=4.79 min; MSCalcd.: 383.7; MS Found: 384.7[M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ1.53-1.81 (m, 2H), 1.86 (s, 3H), 1.89-2.01 (m, 4H), 2.43 (s, 3H),3.41-3.45 (m, 1H), 3.51-3.55 (m, 1H), 6.76 (d, J=6.8 Hz, 1H), 7.04 (dd,J₁=4.4 Hz, J₂=7.2 Hz, 1H), 7.22-7.25 (m, 1H), 7.40 (d, J=7.6 Hz, 1H),7.53 (dd, J₁=4.8 Hz, J₁=7.6 Hz, 1H), 7.62 (d, J=9.2 Hz, 1H), 7.68 (s,1H), 8.01-8.03 (m, 1H), 8.48 (d, J=3.2 Hz, 1H), 8.79 (dd, J₁=1.2 Hz,J₂=5.2 Hz, 1H), 8.86 (d, J=1.6 Hz, 1H).

The Synthesis of X4-119-1

To the solution of X4-101-1 (500 mg, 1.3 mmol) in TEA and THF (20 mL,1:1) were added CuI (12 mg, 0.07 mmol) and PdC12(PPh₃)₂ (92 mg, 0.13mmol). The resulting mixture was heated to 60° C. and stirred for 2hours. After cooled to room temperature, the solid suspension wasfiltered through Celite; the filtrate was concentrated in vacuum and theresidue was purified by flash chromatography to give X4-119-1 (350 mg,66% yield) as off-white solid. LCMS (Agilent LCMS 1200-6120, Column:Waters X-Bridge C18 (30 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.;Flow Rate: 2.0 mL/min; Mobile Phase: from 90% [water+10 mM NH₄HCO₃] and10% [CH₃CN] to 5% [water+10 mM NH₄HCO₃] and 95% [CH₃CN] in 0.5 min, thenunder this condition for 1.5 min, finally changed to 90% [water+10 mMNH₄HCO₃] and 10% [CH₃CN] in 0.1 min and under this condition for 0.7min.); Purity: 90.26%; Rt=0.98 min; MS Calcd.: 421.2; MS Found:422.2[M+H]⁺.

The Synthesis of X4-119-2

To a solution of X4-119-1 (350 mg, 0.83 mmol) in EtOH (20 mL) was addedPd(OH)₂ (20% on carbon) (84 mg, 0.12 mmol) and the mixture was stirredat room temperature for 12 hours under hydrogen atmosphere. Theresulting mixture was filtered through celite and the filtrate wasconcentrated in vacuum, which was purified by column chromatography togive X4-119-2 (130 mg, 37% yield) as off-white syrup. LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 1.6 min, then under this condition for 1.4 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 0.7 min.); Purity: 96.78%; Rt=1.60 min; MS Calcd.:425.2; MS Found: 426.2 [M+H]⁺.

The Synthesis of I-7

Following general procedure A, I-7 (20 mg, 16% yield) was obtained asoff-white solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:100.00%; Rt=1.73 min; MS Calcd.: 411.2; MS Found: 412.4[M+H]⁺. HPLC(Agilent LCMS 1200, Column: Waters X-Bridge C18 (150 mm*4.6 mm*3.5 μm);Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from95% [water+5% TFA] and 5% [CH₃CN] to 0% [water+5% TFA] and 100%[CH₃CN+5% TFA] in 10 min, then under this condition for 5 min, finallychanged to 95% [water+5% TFA] and 5% [CH₃CN] in 0.1 min and under thiscondition for 5 min.); Purity: 90.39%. Rt=4.54 min. ¹H NMR (400 MHz,CDCl₃) δ: 8.60 (1H, dd, J₁=4.8 Hz, J₂=0.8 Hz), 8.50 (1H, br s), 7.74(1H, s), 7.61 (1H, tt, J₁=7.6 Hz, J₂=1.6 Hz), 7.47 (1H, d, J=8.8 Hz),7.42 (1H, d, J=7.2 Hz), 7.19-7.04 (4H, m), 6.58 (1H, d, J=6.8 Hz), 3.57(1H, dd, J₁=11.2 Hz, J₂=2.4 Hz), 3.50-3.45 (1H, m), 3.38-3.27 (4H, m),2.48 (3H, s), 2.07-1.94 (4H, m), 1.90 (3H, s), 1.74-1.61 (2H, m).

Example 5: Synthesis of I-8

The Synthesis of X4-120-1

Following general procedure C, X4-120-1 (351 mg, 43% yield) was obtainedas a brown solid, which was used in the next step without furtherpurification. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min). Purity:88.46%; Rt=1.30 min; MS Calcd.: 201.0; MS Found: 202.7 [M+H]⁺.

The Synthesis of I-8

Following general procedure D, I-8 (18 mg, 22% yield) was obtained aswhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:99.17%; Rt=1.65 min; MS Calcd.: 397.2; MS Found: 398.7 [M+H]⁺. HPLC(Agilent HPLC 1200; Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 93.21%; Rt=4.19 min.¹H NMR (400 MHz, CDCl₃) δ 1.60-1.71 (m, 2H), 1.90-2.02 (m, 4H), 2.56 (s,3H), 2.68-2.81 (m, 4H), 3.99 (t, J=6.8 Hz, 1H), 4.10-4.13 (m, 1H), 6.56(d, J=8.0 Hz, 1H), 6.73 (t, J=6.8 Hz, 1H), 6.84-6.87 (m, 1H), 7.05-7.12(m, 2H), 7.28-7.29 (m, 1H), 7.43 (d, J=8.0 Hz, 1H), 7.56 (d, J=9.2 Hz,1H), 7.61 (s, 1H), 8.04 (d, J=6.8 Hz, 1H), 8.19 (d, J=4.0 Hz, 1H), 8.48(d, J=4.0 Hz, 1H).

Example 6: Synthesis of I-9

The Synthesis of X4-121-1

Following general procedure C, X4-121-1 (115 mg, 71% yield) was obtainedas brown oil, which was used in the next step without furtherpurification. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:44.91%; Rt=1.11 min; MS Calcd.: 202.0; MS Found: 203.7 [M+H]⁺.

The Synthesis of I-9

Following general procedure B, I-9 (12 mg, 15% yield) was obtained asoff-white solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:98.46%; Rt=1.57 min; MS Calcd.: 398.2; MS Found: 399.7 [M+H]⁺. HPLC(Agilent HPLC 1200; Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 96.51%; Rt=4.50 min.¹H NMR (400 MHz, CDCl₃) δ 1.61-1.70 (m, 2H), 1.94-2.06 (m, 4H), 2.51(br, 1H), 2.56 (s, 3H), 2.70-2.75 (m, 2H), 2.79-2.83 (m, 1H), 3.97-4.01(m, 1H), 4.12 (dd, J₁=2.8 Hz, J₂=11.2 Hz, 1H), 6.73-6.77 (m, 1H),7.07-7.15 (m, 2H), 7.44 (d, J=7.6 Hz, 1H), 7.58 (d, J=8.8 Hz, 1H), 7.62(s, 1H), 7.92 (d, J=1.2 Hz, 1H), 8.06 (d, J=6.4 Hz, 1H), 8.12-8.14 (m,2H), 8.49 (d, J=4.0 Hz, 1H).

Example 7: Synthesis of I-10

The Synthesis of I-10

Following general procedure B, I-10 (18 mg, 23% yield) was obtained as awhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:93.04%; Rt=1.63 min; MS Calcd.: 386.2; MS Found: 387.7[M+H]⁺. HPLC(Agilent HPLC 1200; Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 95.05%; Rt=4.95 min.¹H NMR (400 MHz, CDCl₃) δ 1.59-1.66 (m, 2H), 1.93-2.08 (m, 4H), 2.51 (s,3H), 2.63-2.69 (m, 1H), 2.87-2.94 (m, 1H), 3.40 (br, 1H), 3.71-3.75 (m,1H), 3.88 (dd, J₁=2.8 Hz, J₂=11.2 Hz, 1H), 4.04 (dd, J₁=2.4 Hz, J₂=11.2Hz, 1H), 5.96 (t, J=2.0 Hz, 1H), 6.73-6.78 (m, 2H), 7.09-7.16 (m, 3H),7.45-7.46 (m, 2H), 7.56 (d, J=8.8 Hz, 1H), 8.03 (d, J=6.8 Hz, 1H), 8.52(d, J=3.6 Hz, 1H).

Example 8: Synthesis of I-11

The Synthesis of I-11

A mixture of I-1 (120 mg, 0.3 mmol) and acetic acid (0.5 mL) in 37%formaldehyde aq. (10 mL) was stirred at 50° C. for 24 hours, then 37%formaldehyde aq. (5 mL) was added, and the mixture was stirred at 50° C.for another 48 hours. After reaction was completed, the suspension wasadjust to pH 8 with sat. sodium carbonate aq. and extracted with DCM (20mL). The organics layer was concentrated by vacuum and the residue waspurified by prep-HPLC to give I-11 (90 mg, 70% yield) as white solid.LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; MobilePhase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mMNH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under this condition for 1.4min, finally changed to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1min and under this condition for 0.7 min.); Purity: 99.25%; Rt=1.54 min;MS Calcd.: 434.3; MS Found: 435.3 [M+H]⁺. HPLC (Agilent HPLC 1200,Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); Column Temperature: 40°C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95% [water+0.05% TFA] and5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and 100% [CH₃CN+0.05% TFA]in 10 min, then under this condition for 5 min, finally changed to 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1 min and under thiscondition for 5 min.); Purity: 95.82%; Rt=4.28 min. ¹H NMR (400 MHz,CD₃OD) 1.81-1.60 (m, 2H), 1.85 (s, 3H), 1.97-2.01 (m, 2H), 2.10-2.22 (m,2H), 2.45 (s, 6H), 2.48-2.62 (m, 4H), 2.95-3.11 (m, 4H), 3.45-3.50 (m,1H), 3.62-3.70 (m, 2H), 5.32 (d, J=13.6 Hz, 1H), 5.71 (br, 1H),6.70-6.72 (m, 1H), 7.19 (dd, J₁=4.4 Hz, J₂=4.8 Hz, 1H), 7.28-7.35 (m,1H), 7.60 (d, J=7.6 Hz, 1H), 8.39-8.46 (m, 1H).

Example 9: Synthesis of I-12

The Synthesis of I-12

To the solution of I-11 (50 mg, 0.11 mmol) in DCM (5 mL) was addedthionyl chloride (20 mg, 0.17 mmol) at 0° C. under argon atmosphere,then the mixture was stirred at 0° C. for 4 hours. After I-11 has beenconverted completely, the excess of thionyl chloride was removed invacuum; the residue was dissolved in MeOH (2 mL) and stirred at 0° C.for another 2 hours. The mixture was neutralized with NH₃/MeOH andconcentrated in vacuum. The residue was purified by columnchromatography to give I-12 (10 mg, 20% yield) as white solid. LCMS(Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase:from 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [(total 10 mMAcONH₄) H₂O/ACN=1/9 (v/v)] to 10% [(total 10 mM AcONH₄) H₂O/ACN=9/1(v/v)] and 90% [(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 1.6 min, thenunder this condition for 2.4 min, finally changed to 90% [(total 10 mMAcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/ACN=1/9(v/v)] in 0.1 min and under this condition for 0.7 min.); Purity:96.30%; Rt=2.00 min; MS Calcd.: 448.3; MS Found: 449.3[M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 89.34%; Rt=4.56 min.¹H NMR (400 MHz, CD₃OD) 1.20 (s, 3H), 1.55-1.65 (m, 2H), 1.70 (s, 3H),1.84-1.92 (m, 2H), 1.93-2.05 (m, 2H), 2.32 (s, 3H), 2.38-2.57 (m, 4H),2.83-2.94 (m, 4H), 3.11-3.16 (m, 2H), 3.41 (s, 3H), 5.13 (dd, J₁=9.2 Hz,J₂=61.2 Hz, 1H), 6.68 (d, J=6.8 Hz, 1H), 7.12 (d, J=5.6 Hz, 1H),7.21-7.25 (m, 2H), 7.52 (d, J=7.2 Hz, 1H), 8.23 (d, J=2.0 Hz, 1H).

Example 10: Synthesis of I-13, I-14, and I-15

The Synthesis of I-13

Following general procedure I, I-13 (9 mg, 26% yield) was obtained asoff-white solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:98.70%; Rt=1.77 min; MS Calcd.: 418.3; MS Found: 419.4 [M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 90.08%; Rt=4.50 min;MS Calcd.: 418.3; MS Found: 419.4 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.51(d, J=3.6 Hz, 1H), 7.62 (s, 1H), 7.42 (d, J=6.8 Hz, 1H), 7.32 (d, J=10.0Hz, 1H), 7.11-7.15 (m, 1H), 7.05 (dd, J=7.6 Hz, J=4.8 Hz, 1H), 6.28 (d,J=7.2 Hz, 1H), 3.57 (d, J=9.2 Hz, 1H), 3.45-3.49 (m, 1H), 3.18 (s, 4H),2.73 (s, 4H), 2.55-2.61 (m, 2H), 2.48 (s, 3H), 1.92-2.05 (m, 4H), 1.90(s, 3H), 1.62-1.68 (m, 2H), 1.78 (t, 3H).

The Synthesis of I-14

A mixture of I-4 (33.0 mg, 0.085 mmol), X4-189-R (21.7 mg, 0.093 mmol)and DIPEA (12.1 mg, 0.094 mmol) in THF (4 mL) was stirred at 70° C. for4 hours. The mixture was cooled to room temperature, concentrated anddiluted with sat. NaHCO₃aq. (4 mL). The aqueous layer was extracted withDCM three times. The combined organic layers were washed with brine,dried over Na₂SO₄ and filtered. The filtrate was concentrated in vacuumand the residue was purified by prep-HPLC to give I-14 (8 mg, 20% yield)as off-white solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min.); Purity: 91.57%; Rt=1.96 min; MS Calcd.: 472.3; MS Found: 473.4[M+H]⁺. HPLC (Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6mm*5.0 μm); Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; MobilePhase: from 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0%[water+0.05% TFA] and 100% [CH₃CN+0.05% TFA] in 10 min, then under thiscondition for 5 min, finally changed to 95% [water+0.05% TFA] and 5%[CH₃CN+0.05% TFA] in 0.1 min and under this condition for 5 min.);Purity: 89.82%; Rt=6.11 min; MS Calcd.: 472.7; MS Found: 473.4 [M+H]⁺.¹H NMR (400 MHz, CDCl₃) δ 8.50-8.52 (m, 1H), 7.61 (s, 1H), 7.43 (d,J=6.8 Hz, 1H), 7.33 (d, J=8.8 Hz, 1H), 7.14 (dd, J=8.4 Hz, J=7.2 Hz,1H), 7.04-7.07 (m, 1H), 6.27 (d, J=6.4 Hz, 1H), 3.55-3.58 (m, 1H),3.45-3.49 (m, 1H), 3.09-3.17 (m, 6H), 2.96-2.98 (m, 4H), 2.47 (s, 3H),1.92-2.05 (m, 4H), 1.90 (s, 3H), 1.69-1.75 (m, 2H).

The Synthesis of I-15

Following general procedure I, I-15 (9 mg, 25% yield) was obtained asoff-white solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:100.00%; Rt=1.88 min; MS Calcd.: 432.3; MS Found: 433.3 [M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 90.73%; Rt=4.52 min;MS Calcd.: 432.7; MS Found: 433.3 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.51(d, J=4.0 Hz, 1H), 7.63 (s, 1H), 7.42 (d, J=6.8 Hz, 1H), 7.31 (d, J=8.8Hz, 1H), 7.11-7.15 (m, 1H), 7.05 (dd, J=7.6 Hz, J=4.8 Hz, 1H), 6.27 (d,J=6.8 Hz, 1H), 3.57 (d, J=10.0 Hz, 1H), 3.45-3.50 (m, 1H), 3.16 (m, 4H),2.80-2.84 (s, 5H), 2.48 (s, 3H), 1.92-2.09 (m, 4H), 1.91 (s, 3H),1.59-1.67 (m, 2H), 1.15 (d, J=6.4 Hz, 6H).

Example 11: Synthesis of I-16

The Synthesis of I-16

Following general procedure D, I-16 (23 mg, 17% yield) was obtained aswhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:98.43%; Rt=1.77 min; MS Calcd.: 448.2; MS Found: 449.2 [M+H]⁺. HPLC(Agilent HPLC 1200; Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 94.25%; Rt=5.10 min.¹H NMR (400 MHz, CDCl₃) δ 1.57-1.68 (m, 2H), 1.92-1.96 (m, 2H),2.12-2.19 (m, 2H), 2.26 (s, 3H), 3.40-3.44 (m, 1H), 3.58-3.62 (m, 1H),3.74-3.77 (m, 1H), 3.85-3.89 (m, 1H), 6.33-6.34 (m, 1H), 6.63-6.69 (m,2H), 6.79-6.87 (m, 2H), 7.03-7.09 (m, 2H), 7.17-7.18 (m, 1H), 7.24 (d,J=7.6 Hz, 1H), 7.36 (m, 1H), 7.43 (d, J=8.8 Hz, 1H), 7.60 (m, 1H),7.88-7.92 (m, 2H), 8.25-8.26 (m, 1H).

Example 12: Synthesis of I-17

The Synthesis of I-17

Following general procedure K, I-17 (180 mg, 84% yield) was obtained aswhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:96.10%; Rt=1.89 min; MS Calcd.: 482.2; MS Found: 483.3[M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 95.40%; Rt=5.00 min.¹H NMR (400 MHz, MeOD) 1.19-1.52 (m, 2H), 1.54-1.66 (m, 3H), 1.70 (s,3H), 1.85-2.01 (m, 2H), 2.03-2.18 (m, 2H), 2.31 (s, 3H), 2.55-2.63 (m,4H), 2.77-2.86 (m, 4H), 3.50 (br, 2H), 6.55 (dd, J₁=1.6 Hz, J₂=1.6 Hz,1H), 7.10 (dd, J₁=4.8 Hz, J₂=5.2 Hz, 1H), 7.18-7.25 (m, 2H), 7.49 (d,J=8.0 Hz, 1H), 8.18 (d, J=4.8 Hz, 1H).

Example 13: Synthesis of I-18

The Synthesis of I-18

Following general procedure K, I-18 (15 mg, 23% yield) was obtained aswhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0ml/min; Mobile Phase: from 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)]and 10% [(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] to 10% [(total 10 mMAcONH₄) H₂O/ACN=9/1 (v/v)] and 90% [(total 10 mM AcONH₄) H₂O/ACN=1/9(v/v)] in 1.6 min, then under this condition for 2.4 min, finallychanged to 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [(total10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 0.1 min and under this condition for0.7 min.); Purity: 94.68%; Rt=2.02 min; MS Calcd.: 438.2; MS Found:439.2[M+H]⁺. HPLC (Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6mm*5.0 μm); Column Temperature: 40° C.; Flow Rate: 1.0 ml/min; MobilePhase: from 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [total10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] to 15% [total 10 mM AcONH₄) H₂O/ACN=9/1(v/v)] and 85% [total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 5 min, thenunder this condition for 10 min, finally changed to 90% [(total 10 mMAcONH₄) H₂O/ACN=9/1 (v/v)] and 10% [total 10 mM AcONH₄) H₂O/ACN=1/9(v/v)] in 0.1 min and under this condition for 5 min.); Purity: 100%;Rt=6.60 min. ¹H NMR (400 MHz, CD₃OD) 1.18-1.55 (m, 2H), 1.58-1.66 (m,3H), 1.70 (s, 3H), 1.83-2.01 (m, 2H), 2.05-2.18 (m, 2H), 2.30 (s, 3H),2.44-2.65 (m, 4H), 2.79-2.87 (m, 4H), 3.37-3.58 (m, 2H), 6.50 (dd,J₁=2.8 Hz, J₂=5.6 Hz, 1H), 7.10 (dd, J₁=4.8 Hz, J₂=7.6 Hz, 1H),7.16-7.22 (m, 2H), 7.49 (d, J₁=7.2 Hz, 1H), 8.18 (d, J₁=4.0 Hz, 1H).

Example 14: Synthesis of I-19

The Synthesis of I-19

Following general procedure K, I-19 (11 mg, 31% yield) was obtained asoff-white solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0ml/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:96.94%; Rt=2.025 min; MS Calcd.: 496.7; MS Found: 497.7 [M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 ml/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 93.41%; Rt=5.15 min;MS Calcd.: 496.7; MS Found: 497.7 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.48(s, 1H), 7.42-7.37 (m, 2H), 7.12 (dd, J=8.8 Hz, 7.2 Hz, 1H), 7.05 (dd,J=7.6 Hz, 4.8 Hz, 1H), 6.41 (dd, J=7.2 Hz, 0.8 Hz, 1H), 3.61 (dd, J=11.2Hz, 2.4 Hz, 2H), 3.33-3.27 (m, 2H), 2.97-2.85 (m, 4H), 2.60-2.51 (m,4H), 2.46 (s, 3H), 2.19-1.92 (m, 4H), 1.78 (s, 3H), 1.74-1.54 (m, 2H),1.15 (t, 3H).

Example 15: Synthesis of I-20

The Preparation of I-20

Following general procedure K, I-20 (12 mg, 29% yield) was obtained asoff-white solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0ml/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.); Purity:96.27%; Rt=2.140 min; MS Calcd.: 510.7; MS Found: 511.7 [M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 ml/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min.); Purity: 96.35%; Rt=5.268 min;MS Calcd.: 510.7; MS Found: 511.7 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.47(s, 1H), 7.42-7.37 (m, 2H), 7.14-7.10 (m, 1H), 7.05 (dd, J=7.6 Hz, 4.8Hz, 1H), 6.40 (dd, J=7.2 Hz, 0.8 Hz, 1H), 3.61 (d, J=9.6 Hz, 2H),3.34-3.28 (m, 2H), 2.93-2.83 (m, 4H), 2.79-2.69 (m, 3H), 2.47 (s, 3H),2.23-1.92 (m, 4H), 1.79 (s, 3H), 1.74-1.55 (m, 2H), 1.13 (d, J=6.4 Hz,6H).

Example 16: Synthesis of I-21

The synthesis of I-21:

To the solution of I-11 (50 mg, 0.12 mmol) in DCM (5 ml) was addedthionyl chloride (20 mg, 0.17 mmol) at 0° C. under argon atmosphere,then the mixture was stirred at 0° C. for 4 hours. After I-11 wasconsumed completed, the excess thionyl chloride was concentrated byvacuum, the residue was dissolved in ethanol (2 mL) and stirred at 0° C.for another 2 hours. The suspension was diluted with DCM (20 ml) andsaturated sodium bicarbonate (6 ml), the separated organics wasconcentrated by vacuum and purified by prep-HPLC to give I-21 (15 mg,28% yield) as white solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄)H₂O/ACN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/ACN=100/900(v/v)] to 10% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 90% [(total10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 1.6 min, then under this conditionfor 2.4 min, finally changed to 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1(v/v)] and 10% [(total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 0.1 min andunder this condition for 0.7 min.); Purity: 95.10%; Rt=2.08 min; MSCalcd.: 462.3; MS Found: 463.3[M+H]⁺. HPLC (Agilent HPLC 1200, Column:L-column2 ODS (150 mm*4.6 mm*5.0 μm); Column Temperature: 40° C.; FlowRate: 1.0 mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄)H₂O/ACN=9/1 (v/v)] and 10% [total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] to15% [total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 85% [total 10 mM AcONH₄)H₂O/ACN=1/9 (v/v)] in 5 min, then under this condition for 10 min,finally changed to 90% [(total 10 mM AcONH₄) H₂O/ACN=9/1 (v/v)] and 10%[total 10 mM AcONH₄) H₂O/ACN=1/9 (v/v)] in 0.1 min and under thiscondition for 5 min.); Purity: 100%; Rt=6.50 min. 41 NMR (400 MHz, MeOD)1.21-1.24 (m, 3H), 1.63-1.90 (m, 6H), 1.98-2.19 (m, 4H), 2.43 (s, 3H),2.46-2.56 (m, 2H), 2.60-2.74 (m, 2H), 2.94-3.05 (m, 4H), 3.21-3.28 (m,2H), 3.50-3.64 (m, 2H), 3.73-3.79 (m, 2H), 5.16-5.27 (m, 1H), 5.37-5.42(m, 1H), 5.37-5.42 (m, 1H), 6.79 (d, J=7.6 Hz, 1H), 7.23 (dd, J₁=4.8 Hz,J₁=7.6 Hz, 1H), 7.31-7.38 (m, 2H), 7.62 (d, J₁=7.6 Hz, 1H), 8.34 (s,1H).

Example 17: Synthesis of I-34

The Synthesis of Rac-X4-216-2

A mixture of X4-216-1 (10.0 g, 58.0 mmol) and ethyl bromopyruvate (63.9g, 12.9 mmol) in EtOH (60 mL) was stirred at reflux overnight. Then themixture was returned to room temperature, and the solids were collectedby filtration to give X4-216-2 (10.0 g, yield: 64%) as a yellow solid.LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (30 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; MobilePhase: from 90% [water+10 mM NH₄HCO₃] and 10% [CH₃CN] to 5% [water+10 mMNH₄HCO₃] and 95% [CH₃CN] in 0.5 min, then under this condition for 1.5min, finally changed to 90% [water+10 mM NH₄HCO₃] and 10% [CH₃CN] in 0.1min and under this condition for 0.5 min). Purity: 89%, Rt=0.97 min; MSCalcd.: 267.9; MS Found: 268.9 [M+H]⁺.

The Synthesis of X4-216-3

A mixture of X4-216-2 (10.0 g, 37.3 mmol), Selectfluor(1-Chloromethyl-4-fluoro-1,4-diazoniabicyclo[2.2.2]octanebis(tetrafluoroborate)) (26.4 g, 74.6 mmol) and DMAP (4.55 g, 37.3 mmol)in CH₂C12/H₂O (3/1, 40 mL) was stirred at 60° C. overnight. The mixturewas concentrated in vacuum. Water was added to the residue, andextracted with DCM (150 mL×3). The combined organic layers were washedwith brine (50 mL×2), dried over anhydrous Na₂SO₄, filtered andconcentrated in vacuum. The residue was purified by reverse phase columnchromatography to give X4-216-3 (3.00 g, yield: 28%) as a white solid.LCMS (Agilent LCMS 1200-6110, Column: Waters X-Bridge C18 (50 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; MobilePhase: from 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0%[water+0.05% TFA] and 100% [CH₃CN+0.05% TFA] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+0.05% TFA] and 5%[CH₃CN+0.05% TFA] in 0.05 min and under this condition for 0.7 min).Purity: 99%, Rt=1.64 min; MS Calcd.: 286.0; MS Found: 287.1 [M+H]⁺.

The Synthesis of X4-216-4

Following general procedure G, X4-216-4 (1.00 g, yield: 62%) wasobtained as a white solid. LCMS (Agilent LCMS 1200-6110, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+0.05% TFA] and 5%[CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and 100% [CH₃CN+0.05 TFA] in1.6 min, then under this condition for 1.4 min, finally changed to 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.05 min and under thiscondition for 0.7 min). Purity: 96%, Rt=0.58 min; MS Calcd.: 306.1; MSFound: 307.2 [M+H]⁺.

The Synthesis of X4-216-5

To a solution of X4-216-4 (1.00 g, 3.27 mmol) in DCM (20 mL) at −78° C.was added DIBAL-H (6.54 mL, 6.54 mmol) dropwise, and the mixture wasslowly returned to room temperature and stirred for another 2 h. Waterwas added to the residue and the mixture was extracted with DCM (150mL×3). The combined organic layers were washed with brine (50 mL×2),dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum. Theresidue was re-dissolved in DCM (20 mL) and added MnO₂ (2.84 g, 32.7mmol). The mixture was stirred at room temperature overnight andfiltered through Celite. The filtrate was concentrated in vacuum to giveX4-013-5 (700 mg, yield: 82%) as a yellow solid. LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (30 mm*4.6 mm*3.5 μm); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 90%[water+10 mM NH₄HCO₃] and 10% [CH₃CN] to 5% [water+10 mM NH₄HCO₃] and95% [CH₃CN] in 0.5 min, then under this condition for 1.5 min, finallychanged to 90% [water+10 mM NH₄HCO₃] and 10% [CH₃CN] in 0.1 min andunder this condition for 0.5 min). Purity: 77%, Rt=0.99 min; MS Calcd.:262.1; MS Found: 263.0 [M+H]⁺.

The Synthesis of X4-216-6

Following general procedure E, X4-216-6 (500 mg, crude, 72%) wasobtained as a yellow foam, which was used in the next step withoutfurther purification. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (30 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 90% [water+10 mM NH₄HCO₃] and 10%[CH₃CN] to 5% [water+10 mM NH₄HCO₃] and 95% [CH₃CN] in 0.5 min, thenunder this condition for 1.5 min, finally changed to 90% [water+10 mMNH₄HCO₃] and 10% [CH₃CN] in 0.1 min and under this condition for 0.5min.). Purity: 71.5%; Rt=1.07 min; MS Calcd.: 302.1; MS Found: 303.1[M+H]⁺.

The Synthesis of X4-216-7

To a solution of X4-216-6 (500 mg, 1.66 mmol) in MeOH (20 ml) was addedL-proline (76.4 mg, 0.664 mmol), 3-methylpicolinaldehyde (223 mg, 1.83mmol) and MeNH₂ aq. (206 mg, 40% w/w, 6.64 mmol). The reaction mixturewas stirred overnight at room temperature and concentrated in vacuum.The residue was purified by prep-TLC (DCM/MeOH=20:1) to give X4-216-7(200 mg, 28%) as a yellow solid. LCMS (Agilent LCMS 1200-6120, Column:Waters X-Bridge C18 (30 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.;Flow Rate: 2.0 mL/min; Mobile Phase: from 90% [water+10 mM NH₄HCO₃] and10% [CH₃CN] to 5% [water+10 mM NH₄HCO₃] and 95% [CH₃CN] in 0.5 min, thenunder this condition for 1.5 min, finally changed to 90% [water+10 mMNH₄HCO₃] and 10% [CH₃CN] in 0.1 min and under this condition for 0.5min.). Purity: 59.4%. Rt=1.13 min; MS Calcd.: 436.2; MS Found: 437.0[M+H]⁺.

Synthesis of I-34

Following general procedure A, I-34 (7.70 mg, 7%) was obtained as abrown solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄) water/CH₃CN=900/100(v/v)] and 10% [(total 10 mM AcONH₄) water/CH₃CN=100/900 (v/v)] to 10%[(total 10 mM AcONH₄) water/CH₃CN=900/100 (v/v)] and 90% [(total 10 mMAcONH₄) water/CH₃CN=100/900 (v/v)] in 1.6 min, then under this conditionfor 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)water/CH₃CN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄)water/CH₃CN=100/900 (v/v)] in 0.1 min and under this condition for 0.7min) Purity: 91.15%. Rt=1.96 min; MS Calcd.: 422.3; MS Found: 423.2[M+H]⁺. HPLC (Agilent HPLC 1200, Column: Waters X-Bridge C18 (150 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; MobilePhase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mMNH₄HCO₃] and 100% [CH₃CN] in 10 min, then under this condition for 5min, finally changed to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1min and under this condition for 5 min). Purity: 92.8%. Rt=8.85 min. ¹HNMR (400 MHz, CDCl₃) δ 8.47-8.49 (m, 1H), 7.42-7.40 (m, 1H), 7.19-7.16(m, 1H), 7.07-6.99 (m, 2H), 6.18 (d, J=7.2 Hz, 1H), 3.58-3.54 (m, 1H),3.49-3.44 (m, 1H), 3.29-3.24 (m, 2H), 2.98-2.79 (m, 4H), 2.47 (s, 3H),2.39 (s, 3H), 2.25-2.17 (m, 2H), 2.03-1.90 (m, 2H), 1.85 (s, 3H),1.66-1.53 (m, 2H), 1.42-1.22 (m, 2H).

Example 18: Synthesis of I-66

To a solution of X4-019-4 (3.0 g, 1.73 mol) and DME (120 ml) was addedX4-019-4a (5.6 g, 3.03 mol) in one portion at room temperature, thereaction mixture was stirred at 65° C. for 4 hours. After being cooleddown to 10° C., the mixture was stirred for 1 hour and filtered. Afterwashing with DME, The filtered solid was suspended in DME (120 ml) and 2M HCl aq. (120 ml), and stirred overnight at 75° C. After being cooleddown to 10° C., the mixture was neutralized with 3 M NaOH aq. to pH=8and filtered. The filtered cake was washed with water and dried invacuum at 50° C. to give X4-019-5 as an off-white solid (2.81 g, 72%yield). LC-MS (Agilent LCMS 1200-6110, Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 1.6 min, then under this condition for 1.4min, finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA]in 0.05 min and under this condition for 0.7 min). Purity: 94.5%,Rt=1.26 min; MS Calcd.: 223.96; MS Found: 225.1 [M+H]⁺.

Using X4-019-5 and X4-117-B, X4-101-1 was prepared according to GeneralProcedures E and F and methods described herein elsewhere viaintermediate X4-101-0.

The Synthesis of X4-116-A

Following general procedure G, X4-116-A (0.92 g, 24%) was obtained as ayellow solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0ml/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min). Purity:92.35%, Rt=1.81 min; MS Calcd.: 504.7, MS Found: 505.7 [M+1]⁺.

The Synthesis of X4-116-B

Following general procedure A, X4-116-B (140 mg, 16%) was obtained as alight yellow solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 ml/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min). Purity: 93.71%, Rt=1.99 min; MS Calcd.: 490.7, MS Found: 491.7[M+1]⁺.

The Synthesis of I-4

Following general procedure J, I-4 (110 mg, 99%) was obtained as a lightyellow solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0ml/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min). Purity:99.80%, Rt=1.48 min; MS Calcd.: 390.7; MS Found: 391.7 [M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 ml/min; Mobile Phase: from 95%[water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] to 0% [water+0.05% TFA] and100% [CH₃CN+0.05% TFA] in 10 min, then under this condition for 5 min,finally changed to 95% [water+0.05% TFA] and 5% [CH₃CN+0.05% TFA] in 0.1min and under this condition for 5 min). Purity: 98.78%, Rt=4.362 min;MS Calcd.: 390.7; MS Found: 391.7 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.51(d, J=3.6 Hz, 1H), 7.63 (s, 1H), 7.43 (d, J=7.2 Hz, 1H), 7.33 (d, J=8.8Hz, 1H), 7.16-7.12 (m, 1H), 7.07-7.04 (m, 1H), 6.27 (d, J=7.2 Hz, 1H),3.57 (dd, J=11.2, 2.4 Hz, 1H), 3.47 (dd, J=10.0, 3.6 Hz, 1H), 3.15-3.11(m, 8H), 2.47 (s, 3H), 2.22 (s, 3H), 1.97-2.06 (m, 4H), 1.75-1.61 (m,2H).

The Synthesis of I-66

To a solution of I-4 (350.0 mg, 0.90 mmol), X4-276-1 (626.4 mg, 3.6mmol) in THF (5 mL) and MeOH (5 mL) was added NaBH₃CN (223.2 mg, 3.6mmol) and AcOH (0.05 mL). Then the solution was stirred at 60° C. for 7hours. After TLC indicated the reaction completed, the mixture wasconcentrated, quenched with sat. Na2CO3 aq. (20 mL) and extracted withDCM (30 mL×3). The combined organic layers were washed with brine (10mL), dried over Na₂SO₄ and filtered. The filtrate was concentrated invacuum and the residue was purified by prep-HPLC to give I-66 (170 mg,44%) as off-white solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min.) Purity: 96.39%. Rt=1.94 min; MS Calcd.: 430.3; MS Found: 431.4[M+H]⁺. HPLC (Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6mm*5.0 μm); Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; MobilePhase: from 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 10%[total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] to 15% [total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 85% [total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 5 min, then under this condition for 10 min, finally changedto 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 10% [total 10mM AcONH₄) H₂O/MeCN=100/900 (v/v)] in 0.1 min and under this conditionfor 5 min. Purity: 99.9%, Rt=9.61 min. ¹H NMR (400 MHz, CDCl₃) δ 8.51(d, J=3.6 Hz, 1H), 7.63 (s, 1H), 7.43 (dd, J=7.6, 1.2 Hz, 1H), 7.31 (d,J=8.8 Hz, 1H), 7.14-7.10 (m, 1H), 7.05 (dd, J=7.6, 4.8 Hz, 1H), 6.25(dd, J=7.2, 0.8 Hz, 1H), 3.59-3.56 (m, 1H), 3.50-3.45 (m, 1H), 3.11 (s,4H), 2.90 (s, 4H), 2.48 (s, 3H), 2.09-2.05 (m, 2H), 1.97-1.93 (m, 2H),1.91 (s, 3H), 1.83-1.78 (m, 1H), 1.74-1.61 (m, 2H), 0.57-0.52 (m, 2H),0.51-0.46 (m, 2H).

Example 19: Synthesis of I-76

The Synthesis of X4-295-1

To a solution of I-1 (150 mg, 3.7 mmol) in conc. H2SO₄ (2 ml) was addedconc. nitric acid (1.6 mL, 38 mmol) slowly through syringe at 0° C., andthe mixture was stirred at room temperature for 1 hour. Then thereaction mixture was poured into cold NaHCO₃ aqueous solution andextracted with DCM. The organic layer was concentrated in vacuum and theresidue was purified by prep-TLC (DCM/CH₃OH=8:1) to provide productX4-295-1 (40 mg, 26% yield) as yellow solid. LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm)); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 1.6 min, then under this condition for 1.4 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 0.7 min. Purity: 50.59%. Rt=1.64 min; MS Calcd.:449.2; MS Found: 450.4 [M+H]⁺.

The Synthesis of I-76

To a solution of X4-295-1 (100 mg, 0.22 mmol) in ethyl acetate (10 ml)was added Pd(OH)₂/C (50 mg), and the mixture was stirred at 25° C. for36 hours under H2 atmosphere. Then it was filtered through Celite andthe filtrate was concentrated in vacuum. The residue was purified byprep-HPLC to give I-76 (80 mg, 87% yield) as off-white solid. LCMS(Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5μm)); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase:from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mMNH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under this condition for 1.4min, finally changed to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1min and under this condition for 0.7 min. Purity: 91.83%. Rt=1.66 min;MS Calcd.: 419.6; MS Found: 420.4 [M+H]⁺. HPLC (Agilent HPLC 1200,Column: Waters X-Bridge C18 (150 mm*4.6 mm*3.5 μm)); Column Temperature:40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in10 min, then under this condition for 5 min, finally changed to 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this conditionfor 5 min. Purity: 97.02%. Rt=8.05 min. ¹H NMR (400 MHz, CDCl₃) δ 8.50(d, J=3.6 Hz, 1H), 7.55 (1H, s), 7.62 (dd, J=7.6, 1.2 Hz, 1H), 7.32 (dd,J=7.6, 1.2 Hz, 1H), 6.31 (d, J=7.6 Hz, 1H), 6.15 (d, J=7.6 Hz, 1H), 4.30(brs, 2H), 3.57 (dd, J=10.8, 1.6 Hz, 1H,), 3.42 (dd, J=10.8, 1.6 Hz,1H,), 3.05 (s, 4H), 2.66 (s, 4H), 2.44-2.41 (m, 6H), 2.03-1.87 (m, 4H),1.73-1.59 (m, 5H).

Example 20: Synthesis of I-79

The Synthesis of X4-308-A-2

To a solution of X4-308-A-3 (4.00 g, 18.52 mmol) and 51 (4.10 g, 27.71mmol) in the solvent of toluene (36 mL) and water (4 ml) was added K₃PO₄(11.80 g, 55.59 mmol), Pd(dppf)C12-DCM (1.51 g, 1.85 mmol) and PCy₃(0.52 g, 1.85 mmol). The mixture was stirred at room temperature for 28hours and filtered; the filtrate was diluted with water, extracted withEA and concentrated to give a crude product which was purified by CC toafford X4-308-A-2 (703 mg, 21.43%) as yellow oil. LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 1.6 min, then under this condition for 1.4 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 0.7 min). Purity: 82.76%. Rt=1.51 min; MS Calcd.:177.1; MS Found: 178.3 [M+H]⁺.

The Synthesis of X4-308-A-1

To a solution of X4-308-A-2 (175 mg, 0.99 mmol) in THF (3.5 mL) wasadded NaBH₄ (150 mg, 3.96 mmol). The mixture was stirred overnight at70° C., cooled to room temperature, quenched with cooled water,extracted with EA and concentrated to give X4-308-A-1 (120 mg, 81.45%)as yellow oil. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min). Purity:89.49%. Rt=1.38 min; MS Calcd.: 149.1; MS Found: 150.3 [M+H]⁺.

The Synthesis of X4-308-A

To a solution of X4-308-A-1 (550 mg, 3.69 mmol) in DCM (3 ml) was addedDMP (1.73 g, 4.08 mmol). Then the reaction mixture was stirred at roomtemperature for 4 hours. After TLC indicated the reaction completed, thereaction mixture was quenched with NaHCO₃aq. and extracted with DCM. Theorganic layer was washed with brine, dried over Na₂SO₄ and concentratedto give the crude product, which was purified with CC to give X4-308-A(210 mg, 38.70% yield) as yellow oil. LCMS (Agilent LCMS 1200-6120,Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in1.6 min, then under this condition for 1.4 min, finally changed to 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this conditionfor 0.7 min). Purity: 98.67%. Rt=1.51 min; MS Calcd.: 147.1; MS Found:148.3 [M+H]⁺.

The Synthesis of X4-019-6

To a solution of X4-019-5a (1.7 g, 16.7 mmol) and THF (10 ml) was addedn-BuLi (5.6 ml, 14.1 mmol, 2.5 M in hexanes) dropwise at −20° C. Afterstirred at −20° C. for 20 min, the mixture was added slurry of X4-019-5(1.5 g, 6.7 mmol) in THF (30 ml) dropwise at −20° C. and stirred at −10°C. for 7 hours. After quenched with sat NH₄C1 aq. (pH=8), the mixturewas extracted with DCM/i-PrOH (10/1). The organic layer was washed withsat NaHCO₃aq., dried over Na₂SO₄, filtered and concentrated in vacuum.The residue was purified by column chromatography to give productX4-019-6 (785 mg, 48%) as yellow oil. LC-MS (Agilent LCMS 1200-6120,Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in1.6 min, then under this condition for 1.4 min, finally changed to 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this conditionfor 0.7 min). Purity: 94.5%, Rt=1.31 min; MS Calcd.: 224.13; MS Found:245.1 [M+H]⁺.

The Synthesis of X4-014-H

Following general procedure E, X4-014-H (1.9 g, 41%) was obtained as ayellow foam. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min). Purity:69.13%; Rt=1.38 min; MS Calcd.: 284.4; MS Found: 285.4 [M+H]⁺.

The Synthesis of X4-308-1

X4-308-1 (93 mg, 25.86%) was synthesized by reaction of X4-308-A withX4-014-H in the presence of L-proline and methylamine aq. as describedabove to provide the desired product as a yellow solid. LCMS (AgilentLCMS 1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm);Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from90% [(total 10 mM AcONH₄) water/CH₃CN=900/100 (v/v)] and 10% [(total 10mM AcONH₄) water/CH₃CN=100/900 (v/v)] to 10% [(total 10 mM AcONH₄)water/CH₃CN=900/100 (v/v)] and 90% [(total 10 mM AcONH₄)water/CH₃CN=100/900 (v/v)] in 1.6 min, then under this condition for 2.4min, finally changed to 90% [(total 10 mM AcONH₄) water/CH₃CN=900/100(v/v)] and 10% [(total 10 mM AcONH₄) water/CH₃CN=100/900 (v/v)] in 0.1min and under this condition for 0.7 min.) Purity: 38.95%. Rt=1.66 min;MS Calcd.: 444.3; MS Found: 445.3 [M+H]⁺.

The Synthesis of I-79

following General Procedure A, I-79 (12 mg, 13.32%) was synthesized as alight yellow solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min). Purity: 100.00%. Rt=1.756 min; MS Calcd.: 430.3; MS Found: 431.4[M+H]⁺. HPLC (Agilent HPLC 1200, Column: Waters X-Bridge C18 (150 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; MobilePhase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mMNH₄HCO₃] and 100% [CH₃CN] in 10 min, then under this condition for 5min, finally changed to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1min and under this condition for 5 min). Purity: 98.73%. Rt=8.82 min. ¹HNMR (400 MHz, CDCl₃) δ 8.49 (s, 1H), 7.51 (s, 1H), 7.33-7.27 (m, 2H),7.14 (dd, J=9.0, 7.0 Hz, 1H), 7.06 (dd, J=8.0, 4.8 Hz, 1H), 6.27 (dd,J=7.2, 0.8, 1H), 3.97 (d, J=8.4 Hz, 1H), 3.48 (dd, J=11.6, 2.4 Hz, 1H),3.18-3.14 (m, 4H), 2.77-2.70 (m, 4H), 2.43 (s, 3H), 2.13-2.05 (m, 3H),1.99-1.94 (2H, m), 1.89 (s, 3H), 1.77 (d, J=9.2 Hz, 1H), 1.67-1.62 (m,2H), 1.07-1.00 (m, 2H), 0.72-0.69 (m, 1H), 0.68-0.51 (m, 1H).

Example 21: Synthesis of I-146

The Synthesis of X4-438-1

A mixture of X4-014-H (2.5 g, 8.8 mmol), 3-methylpicolinaldehyde (1.1 g,8.8 mmol), K₂CO₃ (1.8 g, 13.2 mmol) in toluene (100 ml)/EtOH (40 ml)/H₂O(20 ml) was stirred at 80° C. overnight. After reaction completed, themixture was cooled down to room temperature and concentrated in vacuum.The residue was purified by flash silica gel pad (3 cm), eluted withEtOAc:petroleum ether=1:1 to DCM/MeOH=100/1 to give 2.3 g of X4-438-1 asyellow foam (yield: 68%), which was used for the next step directly.LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min; MobilePhase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mMNH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under this condition for 1.4min, finally changed to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1min and under this condition for 0.7 min). Purity: 77.36%; Rt=1.59 min;MS Calcd.: 387.2; MS Found: 388.3 [M+H]⁺.

The Synthesis of I-204

To a solution of X4-438-1 (2.3 g, 5.9 mmol) in MeOH (120 ml) was addedconc. NH₃ aq. (5 ml, 20%, 59 mmol) at room temperature and the mixturewas stirred at room temperature overnight. After reaction completed, themixture was concentrated in vacuum and the residue was purified by flashsilica gel pad (3 cm), eluting with DCM/MeOH=60/1 to 30/1 to give 1 g ofI-204 as yellow foam (yield: 42%), which was used for the next stepdirectly. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min;Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0%[water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min). Purity:78.15%; Rt=1.41 min; MS Calcd.: 404.2; MS Found: 405.4 [M+H]⁺.

The Synthesis of I-187

A mixture of I-204 (1 g, 2.5 mmol), KOH (2.8 g, 50 mmol) and N₂H₄.H₂O (5g, 100 mmol) in diethylene glycol (30 ml) was stirred at 80° C. for 2 h;then removed N₂H₄.H₂O in vacuum when heating to 160° C. and stirred at160° C. for 2 h. After the reaction was completed, the mixture wascooled down to room temperature and quenched with H₂O (90 ml)/DCM (120ml). The aqueous layer was extracted with DCM (120 ml×3). The combinedorganic layers were washed with H₂O and brine, dried over Na₂SO₄,filtered and concentrated in vacuum. The residue was purified by CC(eluted with DCM/MeOH) to give 220 mg of (racemic) cis-I-187 (220 mg),400 mg of (racemic) mix of cis/trans-I-187 (400 mg, trans:cis=4:5) and270 mg of (racemic) trans-I-187 (270 mg). Yield=91%. LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 1.6 min, then under this condition for 1.4 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 0.7 min). Cis: Purity: 71.18%; Rt=1.51 min; Trans:Purity: 64.10%; Rt=1.48 min; Mix: Purity: 84.07%; MS Calcd.: 390.3; MSFound: 391.4 [M+H]⁺.

The Synthesis of X4-440-1

To a solution of cis/trans-I-187 (300 mg, 0.77 mmol) in dichloromethane(10 mL) was added triethylamine (233 mg, 2.30 mmol) and di-tert-butyldicarbonate (252 mg, 1.15 mmol), and the mixture was stirred at 40° C.for 4 h. After reaction was completed, the suspension was diluted withwater (10 mL) and dichloromethane (20 mL). The separated organic layerwas concentrated by vacuum and the residue was purified by prep-TLC(eluted with ethyl acetate) to give X4-440-1 (100 mg, 27%) as whitesolid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min;Mobile Phase: from 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] to 10% [(total 10 mMAcONH₄) H₂O/MeCN=900/100 (v/v)] and 90% [(total 10 mM AcONH₄)H₂O/MeCN=100/900 (v/v)] in 1.6 min, then under this condition for 2.4min, finally changed to 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100(v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] in 0.1 minand under this condition for 0.7 min. Purity: 80.70%, Rt=2.10 min; MSCalcd.: 490.3; MS Found: 491.3 [M+H]⁺.

The Synthesis of X4-440-2

Following general procedure K, X4-440-2 (80 mg, 100%) was obtained aswhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100(v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] to 10%[(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 90% [(total 10 mMAcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min, then under this conditionfor 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 0.1 min and under this condition for 0.7 min. Purity: 96.33%,Rt=2.14 min; MS Calcd.: 524.3; MS Found: 525.2 [M+H]⁺.

The Synthesis of I-146

Following general procedure J, I-146 (11 mg, 17%) was obtained as whitesolid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min;Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0%[water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.). Purity:97.67%; Rt=1.70 min; MS Calcd.: 424.2; MS Found: 425.4[M+H]⁺. HPLC(Agilent HPLC 1200, Column: Waters X-Bridge C18 (150 mm*4.6 mm*3.5 μm);Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 10 min, then under this condition for 5 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 5 min.). Purity: 97.60%, Rt=8.21 min; MS Calcd.:424.2; MS Found: 425.4[M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.40 (dd, J=4.8,1.2 Hz, 1H), 7.38 (dd, J=7.2, 1.6 Hz, 1H), 7.32 (dd, J=9.2, 1.2 Hz, 1H),7.08 (dd, J=8.8, 7.2 Hz, 1H), 7.02 (dd, J=8.0, 4.8 Hz, 1H), 6.34 (dd,J=7.2, 0.8 Hz, 1H), 4.25-4.18 (m, 2H), 3.29-3.25 (m, 2H), 2.96-2.78 (m,4H), 2.53-2.46 (m, 2H), 2.38 (s, 6H), 2.16-2.11 (m, 1H), 1.88-1.80 (m,4H), 1.72-1.65 (m, 2H).

Example 22: Synthesis of I-149, I-188, I-189, and I-205

The Synthesis of X4-443-1

To a solution of X4-014-H (2.0 g, 7.03 mmol), X4-014-B (992.6 mg, 7.01mmol) in toluene/EtOH/H₂O (5:2:1, 50 mL) was added K₂CO₃ (1.46 g, 10.56mmol). The mixture was stirred at 80° C. overnight. The reaction mixturewas cooled to room temperature, concentrated in vacuum and the residuewas purified by column chromatography to give X4-443-1 (2.0 g, 70%) asyellow solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min). Purity:79.84%. Rt=1.65 min; MS Calcd.: 407.2; MS Found: 408.2 [M+H]⁺.

The Synthesis of I-205

To a solution of X4-443-1 (2.0 g, 4.9 mmol) in MeOH (120 mL) were addedNH₃ aq. (20%, 4.2 mL, 49 mmol), and the mixture was stirred at roomtemperature overnight. Then it was concentrated in vacuum and theresidue was purified by column chromatography to give I-205 (600 mg, 29%yield, 65% trans; 25% cis) as yellow solid. LCMS (Agilent LCMS1200-6120, Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); ColumnTemperature: 40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 95%[water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 1.6 min, then under this condition for 1.4 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 0.7 min). Purity: 71.43%. Rt=1.44 min, 1.47 min; MSCalcd.: 424.2; MS Found: 425.2 [M+H]⁺.

The Synthesis of I-149, I-188, and I-189

Following general procedure A, I-149 (30 mg, 22% yield) was obtained aslight-yellow solid. The solid was separated by SFC to give I-188 (10 mg)and I-189 (11 mg). LCMS (Agilent LCMS 1200-6120, Column: Waters X-BridgeC18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0ml/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.). Purity:97.24%, Rt=1.60 min; MS Calcd.: 410.2; MS Found: 411.2 [M+H]⁺. HPLC(Agilent HPLC 1200, Column: L-column2 ODS (150 mm*4.6 mm*5.0 μm); ColumnTemperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from 90%[(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 10% [total 10 mMAcONH₄) H₂O/MeCN=100/900 (v/v)] to 15% [total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 85% [total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 5 min, then under this condition for 10 min, finally changedto 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 10% [total 10mM AcONH₄) H₂O/MeCN=100/900 (v/v)] in 0.1 min and under this conditionfor 5 min. Purity: >99.00%, Rt=5.63 min. ¹H NMR (400 MHz, CDCl₃) δ8.59-8.50 (dd, J=4.8, 1.6 Hz, 1H), 7.65-7.63 (m, 1H), 7.51 (s, 1H), 7.32(d, J=8.8 Hz, 1H), 7.17-7.10 (m, 2H), 6.27 (dd, J=7.2, 0.8 Hz, 1H), 4.52(dd, J=11.2, 2.4, 1H), 4.20-4.17 (m, 1H), 3.14 (s, 4H), 2.66 (m, 4H),2.40 (s, 3H), 2.23-2.19 (m, 1H), 2.15-2.11 (m, 1H), 2.02-1.98 (m, 1H),1.89-1.77 (m, 2H), 1.61-1.54 (m, 1H).

Example 23: Synthesis of I-154 and I-206

The Synthesis of X4-449-1

To the solution of X4-027-A (10.0 g, 68.42 mmol) in dichloromethane (60mL)/water (20 mL) was added selectfluor (10.0 g, 68.42 mmol) and4-dimethylaminopyridine (10.0 g, 68.42 mmol) in portions at 0° C., andthe mixture was stirred at room temperature overnight. After reactionwas completed, the suspension was separated and the organics wasconcentrated by vacuum. The residue was purified by silica gel, elutingwith petroleum ether/ethyl acetate 5:1 to give X4-449-1 (6.0 g, 54%) aswhite solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100(v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] to 10%[(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 90% [(total 10 mMAcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min, then under this conditionfor 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 0.1 min and under this condition for 0.7 min. Purity: 63.38%,Rt=0.94 min; MS Calcd.: 164.0; MS Found: 165.2 [M+H]⁺.

The Synthesis of X4-451-1

A mixture of X4-449-1 (1.3 g, 7.92 mmol) and 1-methylpiperazine (1.6 g,15.84 mmol) in acetonitrile (10 mL) was stirred at 80° C. for 1 hour,then diluted with dichloromethane (60 mL) and water (20 mL). Theseparated organic layer was concentrated by vacuum, then purified bysilica gel, eluting with petroleum ether/ethyl acetate 3:1 to giveX4-449-1 (1.5 g, 78%) as pale solid. LCMS (Agilent LCMS 1200-6120,Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 90% [(total 10 mMAcONH₄) H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄)H₂O/MeCN=100/900 (v/v)] to 10% [(total 10 mM AcONH₄) H₂O/MeCN=900/100(v/v)] and 90% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min,then under this condition for 2.4 min, finally changed to 90% [(total 10mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄)H₂O/MeCN=100/900 (v/v)] in 0.1 min and under this condition for 0.7 min.Purity: 71.35%, Rt=1.02 min; MS Calcd.: 244.1; MS Found: 245.3 [M+H]⁺.

The Synthesis of X4-451-2

Following general procedure E, X4-451-2 (580 mg, 33%) was obtained asyellow solid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100(v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] to 10%[(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 90% [(total 10 mMAcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min, then under this conditionfor 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 0.1 min and under this condition for 0.7 min. Purity: 93.40%,Rt=1.27 min; MS Calcd.: 284.2; MS Found: 285.2 [M+H]⁺.

The Synthesis of X4-451-3

A mixture of X4-451-2 (400 mg, 1.41 mmol), potassium carbonate (293 mg,2.12 mmol) and 3-methylpicolinaldehyde (188 mg, 1.55 mmol) in toluene (5mL)/ethanol (2 mL)/water (1 mL) was stirred at 80° C. overnight. Themixture was diluted with dichloromethane (30 mL) and water (10 mL), theseparated organics was concentrated by vacuum and purified by flashsilica gel column by eluting with dichloromethane/methanol 100:1 to giveX4-451-3 (500 mg, 92%) as yellow solid. LCMS (Agilent LCMS 1200-6120,Column: Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 90% [(total 10 mMAcONH₄) H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄)H₂O/MeCN=100/900 (v/v)] to 10% [(total 10 mM AcONH₄) H₂O/MeCN=900/100(v/v)] and 90% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min,then under this condition for 2.4 min, finally changed to 90% [(total 10mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄)H₂O/MeCN=100/900 (v/v)] in 0.1 min and under this condition for 0.7 min.Purity: 75.42%, Rt=1.65 min; MS Calcd.: 387.2; MS Found: 388.2 [M+H]⁺.

The Synthesis of I-206

A mixture of X4-451-3 (500 mg, 1.29 mmol) and ammonium hydroxide (878mg, 12.90 mmol, 25% wt) in menthol (25 mL) was stirred at roomtemperature overnight. After reaction was completed, the solvent wasremoved by vacuum, and the residue was purified by silica gel column byeluting with dichloromethane/methanol 80:1 to give I-206 (100 mg, 19%)as light yellow solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] to 10% [(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 90%[(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min, then underthis condition for 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 0.1 min and under this condition for 0.7 min. Purity: 75.12%,Rt=1.37 min; MS Calcd.: 404.2; MS Found: 405.2 [M+H]⁺.

The Synthesis of I-154

Following general procedure A, I-154 (18 mg, 23%) was obtained as palesolid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (50mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min;Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0%[water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, then under thiscondition for 1.4 min, finally changed to 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] in 0.1 min and under this condition for 0.7 min.). Purity:99.55%; Rt=1.53 min; MS Calcd.: 390.3; MS Found: 391.3[M+H]⁺. HPLC(Agilent HPLC 1200, Column: Waters X-Bridge C18 (150 mm*4.6 mm*3.5 μm);Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 10 min, then under this condition for 5 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 5 min). Purity: 97.86%, Rt=7.07 min. ¹H NMR (400 MHz,CDCl₃) δ 8.40 (dd, J=4.4, 0.8 Hz, 1H), 8.04 (d, J=6.8 Hz, 1H), 7.49 (d,J=9.2 Hz, 1H), 7.37 (dd, J=7.6, 0.8 Hz, 1H), 7.09-7.05 (m, 1H), 7.00(dd, J=8.0, 4.8 Hz, 1H), 6.73-6.69 (m, 1H), 4.29-4.23 (m, 1H), 4.16-4.11(m, 1H), 3.43-3.11 (m, 4H), 3.10-2.72 (m, 4H), 2.41 (s, 3H), 2.36 (s,3H), 2.14-2.10 (m, 1H), 1.90-1.76 (m, 4H), 1.67-1.59 (m, 2H).

Example 24: Synthesis of I-187

Following general procedure A, 30 mg of I-187 was obtained as whitesolid from I-204 (1 g, 2.5 mmol). LCMS (Agilent LCMS 1200-6120, Column:Waters X-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.;Flow Rate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min.). Cis: Purity: 97.37%; Rt=1.51 min; MS Calcd.: 390.3; MS Found:391.4 [M+H]⁺. HPLC (Agilent HPLC 1200, Column: Waters X-Bridge C18 (150mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 1.0 mL/min;Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0%[water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 10 min, then under thiscondition for 5 min, finally changed to 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] in 0.1 min and under this condition for 5 min.). Purity: 90.65%,Rt=6.95 min. ¹H NMR (400 MHz, CDCl₃) δ 8.43 (dd, J=4.8, 1.2 Hz, 1H),7.42 (dd, J=7.2, 1.6 Hz, 1H), 7.31 (d, J=9.2 Hz, 1H), 7.13 (dd, J=9.2,7.2 Hz, 1H), 7.05 (dd, J=7.6, 4.8 Hz, 1H), 6.27 (dd, J=7.2, 1.2 Hz, 1H),4.24-4.21 (m, 1H), 4.18-4.15 (m, 1H), 3.14 (s, 4H), 2.65 (s, 4H), 2.40(s, 6H), 2.25-2.22 (m, 1H), 2.15-2.12 (m, 1H), 1.85-1.76 (m, 3H),1.72-1.65 (m, 2H).

Example 25: Synthesis of I-191

The Synthesis of X4-485-1

A mixture of X4-014-H (1.0 g, 3.52 mmol), potassium carbonate (730 mg,5.28 mmol) and 3-methoxypicolinaldehyde (482 mg, 3.52 mmol) in toluene(25 mL)/ethanol (10 mL)/water (5 mL) was stirred at 80° C. overnight.After the reaction was completed, the mixture was diluted withdichloromethane (50 mL) and water (10 mL), the separated organic layerswere concentrated by vacuum, and purified by a flash silica gel pad (3cm), eluting with dichloromethane/methanol 80:1 to give X4-485-1 (1.0 g,70%) as yellow solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] to 10% [(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 90%[(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min, then underthis condition for 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 0.1 min and under this condition for 0.7 min. Purity: 82.63%,Rt=1.67 min; MS Calcd.: 403.2; MS Found: 404.2 [M+H]⁺.

The Synthesis of X4-485-2

A mixture of X4-485-1 (1.0 g, 2.48 mmol) and ammonium hydroxide (1.7 g,24.78 mmol, 25% wt) in methanol (50 mL) was stirred at room temperatureovernight. After the reaction was completed, the organic layer wasconcentrated and purified by silica gel column by eluting withdichloromethane/methanol 50:1 to give X4-485-2 (600 mg, 58%) as lightyellow oil. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18(50 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0mL/min; Mobile Phase: from 90% [(total 10 mM AcONH₄) H₂O/MeCN=900/100(v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900 (v/v)] to 10%[(total 10 mM AcONH₄) H₂O/MeCN=900/100 (v/v)] and 90% [(total 10 mMAcONH₄) H₂O/MeCN=100/900 (v/v)] in 1.6 min, then under this conditionfor 2.4 min, finally changed to 90% [(total 10 mM AcONH₄)H₂O/MeCN=900/100 (v/v)] and 10% [(total 10 mM AcONH₄) H₂O/MeCN=100/900(v/v)] in 0.1 min and under this condition for 0.7 min. Purity: 82.42%,Rt=1.35 min; MS Calcd.: 420.2; MS Found: 421.3 [M+H]⁺.

The Synthesis of I-191

Following general procedure A, I-191 (impure, 280 mg, 48%) was obtainedas yellow solid, and 140 mg was purified by prep-HPLC to give pure I-191(80 mg). LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (30mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min;Mobile Phase: from 90% [water+10 mM NH₄HCO₃] and 10% [CH₃CN] to 5%[water+10 mM NH₄HCO₃] and 95% [CH₃CN] in 0.5 min, then under thiscondition for 1.5 min, finally changed to 90% [water+10 mM NH₄HCO₃] and10% [CH₃CN] in 0.1 min and under this condition for 0.7 min. Purity:100.00%; Rt=1.08 min; MS Calcd.: 406.2; MS Found: 407.1 [M+H]⁺. HPLC(Agilent HPLC 1200, Column: Waters X-Bridge C18 (150 mm*4.6 mm*3.5 μm);Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; Mobile Phase: from95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mM NH₄HCO₃] and100% [CH₃CN] in 10 min, then under this condition for 5 min, finallychanged to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1 min and underthis condition for 5 min). Purity: 92.84%, Rt=6.85 min; MS Calcd.:406.2; MS Found: 407.4 [M+H]⁺. ¹H NMR (400 MHz, CDCl₃) δ 8.18 (t, J=6.0Hz, 1H). 7.53 (s, 1H), 7.31 (d, J=9.2 Hz, 1H), 7.15-7.11 (m, 3H), 6.26(dd, J=7.2, 0.8 Hz, 1H), 4.46 (dd, J=1.6, 1.2 Hz, 1H), 4.19 (d, J=10.0Hz, 1H), 3.85 (s, 3H), 3.13 (s, 4H), 2.65 (s, 4H), 2.39 (s, 3H),2.24-2.20 (m, 1H), 2.11-2.07 (m, 1H), 1.95-1.92 (m, 1H), 1.86-1.76 (m,3H), 1.70-1.63 (m, 1H).

Example 26: Synthesis of Additional Exemplary Compounds

Additional exemplary compounds were prepared following methodssubstantially similar to those described above and herein. Data forthese compounds are provided below.

TABLE 2 Characterization Data for Additional Exemplary Compounds RtCompound (Min) Rt (Min) No. Chemical Structure M + 1 (LCMS) (HPLC) ¹HNMR (400 MHz) I-25

447.4 1.56 4.45 (CDCl₃) δ 8.42 (s, 1H), 8.35 (d, J = 6.8 Hz, 1H), 7.56(d, J = 9.2 Hz, 1H), 7.41-7.39 (m, 1H), 7.11- 7.03 (m, 2H), 6.77-6.73(m, 1H), 3.58-3.54 (m, 2H), 3.48-3.45 (m, 2H), 3.11 (brs, 2H), 2.56-2.49(m, 4H), 2.47-2.45 (m, 4H), 2.44-2.42 (m, 2H), 2.42-2.41 (m, 2H), 2.30(s, 3H), 2.14-2.04 (m, 2H), 2.03-1.91 (m, 2H), 1.85- 1.81 (m, 3H), 1.78(s, 3H), 1.74-1.59 (m, 2H) I-26

364.4 1.40 6.91 (CDCl₃) δ 8.47 (s, 1H), 7.95 (d, J = 6.8 Hz, 1H), 7.56(d, J = 8.8 Hz, 1H), 7.41 (d, J = 7.2 Hz, 1H), 7.12-7.03 (m, 2H), 6.77-6.74 (m, 1H), 3.73-3.71 (m, 1H), 3.55-3.41 (m, 1H), 3.13 (brs, 2H),2.93- 2.82 (m, 3H), 2.13-2.00 (m, 2H), 1.97-1.82 (m, 4H), 1.76-1.69 (m,5H), 1.63-1.22 (m, 2H). I-27

378.4 1.39 4.42 (CDCl₃) δ 8.44 (s, 1H). 7.97 (d, J = 6.8 Hz, 1H), 7.57(d, J = 8.8 Hz, 1H), 7.41 (d, J = 7.2 Hz, 1H), 7.12-7.03 (m, 2H), 6.78-6.74 (m, 1H), 3.56 (dd, J = 11.6, 2.8 Hz, 1H), 3.47 (dd, J = 11.6, 2.4Hz, 1H), 3.16-3.08 (m, 2H), 2.74- 2.67 (m, 3H), 2.55-2.52 (m, 3H), 2.47(s, 3H), 2.09-1.99 (m, 2H), 1.94- 1.90 (m, 2H), 1.85-1.81 (m, 2H), 1.80(s, 3H), 1.75-1.55 (m, 2H). I-28

392.3 1.59 5.71 (CDCl₃) δ 8.46 (s, 1H), 7.98 (d, J = 6.8 Hz, 1H), 7.57(d, J = 9.2 Hz, 1H), 7.41 (d, J = 6.8 Hz, 1H), 7.12-7.03 (m, 2H), 6.77-6.74 (m, 1H), 3.56 (dd, J = 9.2, 2.8 Hz, 1H), 3.45 (dd, J = 11.6, 2.4Hz, 1H), 3.06 (s, 2H), 2.71 (s, 6H), 2.49 (s, 2H), 2.37-2.34 (m, 2H),2.06-2.02 (m, 2H), 1.94- 1.91 (m, 2H), 1.82-1.78 (m, 6H), 1.60-1.23 (m,2H). I-29

434.3 1.87 8.15 (CDCl₃) δ 8.46 (s, 1H), 8.01 (d, J = 6.8 Hz, 1H), 7.58(d, J = 15.4 Hz, 1H), 7.41 (d, J = 8.0 Hz, 1H), 7.12-7.04 (m, 2H), 6.77-6.73 (m, 1H), 3.75-3.73 (m, 4H), 3.56 (dd, J = 11.2, 2.8 Hz, 1H), 3.45(dd, J = 11.6, 2.4 Hz, 1H), 3.08 (s, 2H), 2.45-2.25 (m, 9H), 2.17-1.95(m, 2H), 1.94-1.85 (m, 2H), 1.83- 1.59 (m, 7H), I-34

423.2 1.96 8.85 (CDCl₃) δ 8.47-8.49 (m, 1H), 7.42-7.40 (m, 1H),7.19-7.16 (m, 1H), 7.07- 6.99 (m, 2H), 6.18 (d, J = 7.2 Hz, 1H),3.58-3.54 (m, 1H), 3.49-3.44 (m, 1H), 3.29-3.24 (m, 2H), 2.98- 2.79 (m,4H), 2.47 (s, 3H), 2.39 (s, 3H), 2.25-2.17 (m, 2H), 2.03-1.90 (m, 2H),1.85 (s, 3H), 1.66-1.53 (m, 2H), 1.42-1.22 (m, 2H). I-37

447.4 2.67 9.97 (CDCl₃) δ 8.43 (d, J = 4.0 Hz, 1H, ), 7.59 (brs, 1H),7.36 (d, J = 7.2 Hz, 1H), 7.24 (d, J = 8.8 Hz, 1H, , 7.06 (t, J = 7.6Hz, 1H), 6.98 (q, J = 7.2 Hz, 1H), 6.21 (d, J = 7.2 Hz, 1H), 3.91 (brs,2H), 3.09 (brs, 4H), 2.75 (brs, 5H), 2.44 (brs, 5H), 1.90-1.55 (m, 6H),1.09 (d, J = 6.4 Hz, 6H), 0.58 (t, J = 6.8 Hz, 3H). I-45

525.3 3.14 11.60 (CDCl₃) δ 8.36 (s, 1H), 7.34-7.31 (m, 2H), 7.06- 6.97(m, 2H), 6.33 (d, J = 6.8 Hz, 1H), 3.99 (brs, 2H), 3.25 (brs, 2H), 2.79(brs, 6H), 2.45 (brs, 3H), 2.14 (brs, 2H), 1.97-1.82 (m, 2H), 1.70-1.65(m, 4H), 1.07 (s, 6H), 0.58 (t, J = 6.8 Hz, 3H). I-46

497.2 2.10 10.07 (CDCl₃) δ 8.37 (d, J = 3.2 Hz, 1H), 7.45-7.32 (m, 2H),7.19-6.97 (m, 2H), 6.34 (d, J = 6.8 Hz, 1H), 4.05 (brs, 2H), 3.23 (brs,2H), 2.90-2.77 (m, 4H), 2.56 (brs, 5H), 2.35 (brs, 5H), 2.04-1.55 (m,6H), 0.59 (t, J = 6.8 Hz, 3H). I-47

511.2 2.78 10.38 (CDCl₃) δ 8.38 (d, J = 3.6 Hz, 1H), 7.33-7.29 (m, 2H),7.18-6.96 (m, 2H), 6.32 (tr, J = 7.2 Hz, 1H), 3.20 (d, J = 6.4 Hz, 2H),3.98 (brs, 2H), 2.87-2.75 (m, 4H), 2.62-2.45 (m, 6H), 2.33 (s, 3H),2.08- 1.62 (m, 6H), 0.62 (d, J = 6.8 Hz, 6H). I-87

435.2 1.48 7.36 (DMSO-d₆) δ 8.41 (d, J = 3.6 Hz, 1H), 7.60 (s, 1H), 7.39(d, J = 7.6 Hz, 1H), 7.26-7.24 (m, 1H), 7.13- 7.01 (m, 2H), 6.24 (d, J =6.8 Hz, 1H), 3.73 (brs, 1H), 3.17-3.04 (m, 4H), 2.43-2.37 (m, 8H), 2.02-1.90 (m, 4H), 1.74-1.69 (m, 2H). I-49

449.4 2.01 7.29 (CDCl₃) δ 8.43 (d, J = 3.2 Hz, 1H), 7.58 (s, 1H),7.38-7.35 (m, 1H), 7.25- 7.20 (m, 1H), 7.11-7.07 (m, 1H), 7.01-6.98 (m,1H), 6.22 (dd, J = 7.2, 0.8 Hz, 1H), 3.87-3.78 (m, 3H), 3.29-3.03 (m,6H), 2.63-2.49 (m, 5H), 2.46- 2.36 (m, 7H), 2.07-1.69 (m, 8H). I-51

433.3 0.20 9.31 (CDCl₃) δ 8.49 (s, 1H), 7.59 (d, J = 7.6 Hz, 2H), 7.32(d, J = 9.2 Hz, 1H), 7.16-7.12 (m, 2H), 6.28 (d, J = 6.4 Hz, 1H), 3.67(d, J = 10.0 Hz, 1H), 3.48-3.46 (m, 1H), 3.16 (d, J = 16.0 Hz, 4H), 2.70(s, 4H), 2.43 (s, 3H), 2.06-1.90 (m, 6H), 1.76 (s, 4H), 1.27 (t, J = 6.8Hz, 6H). I-52

447.3 1.59 7.61 (CDCl₃) δ 8.51 (d, J = 4.4 Hz, 1H), 7.62 (s, 1H),7.44-7.42 (m, 1H), 7.33 (d, J = 8.8 Hz, 1H), 7.14 (dd, J = 8.8, 7.2 Hz,1H), 7.05 (dd, J = 7.6, 4.8 Hz, 1H), 6.30 (dd, J = 7.2, 1.8 Hz, 1H),4.76 (t, J = 6.4 Hz, 2H), 4.69 (t, J = 6.4 Hz, 2H), 3.71-3.65 (m, 1H),3.56 (dd, J = 11.6, 2.4 Hz, 1H), 3.47 (t, 1H), 3.19 (s, 4H), 2.63 (s,4H), 2.47 (s, 3H), 2.08-1.95 (m, 4H), 1.90 (s, 3H), 1.73-1.59 (m, 2H).I-56

477.4 2.66 9.83 (CD₃OD) δ 8.46 (brs, 1H), 7.67-7.66 (m, 1H), 7.40- 7.26(m, 3H), 6.83-6.82 (m, 1H), 5.41-5.36 (m, 1H), 5.21-5.18 (m, 1H),3.78-3.77 (m, 2H), 3.50- 3.40 (m, 2H), 3.15-2.95 (m, 5H), 2.65-2.44 (m,7H), 2.23-1.78 (m, 9H), 1.18-1.31 (m, 7H) I-57

491.4 2.03 4.63 (CDCl₃) δ 8.35 (d, J = 3.2 Hz, 1H). 7.53 (d, J = 6.8 Hz,1H), 7.32-7.30 (m, 1H), 7.23-7.14 (m, 2H), 6.67-6.66 (d, J = 6.8 Hz,1H), 5.20-5.05 (m, 2H), 3.64-3.59 (m, 2H), 3.54- 3.44 (m, 2H), 3.18-3.13(t, 2H), 2.85-2.72 (m, 5H), 2.52-2.46 (m, 5H), 2.02- 1.53 (m, 9H),1.14-1.02 (m, 9H). I-58

489.4 2.09 10.85 (CDCl₃) δ 8.46 (s, 1H), 7.42-7.38 (m, 2H), 7.12- 7.04(m, 2H), 6.49 (d, J = 6.8 Hz, 1H), 5.31 (d, J = 11.2 Hz, 1H), 5.12 (d, J= 8.4 Hz, 1H), 3.71-3.65 (m, 2H), 3.59-3.52 (m, 2H), 3.24 (d, J = 9.6Hz, 2H), 3.07-3.03 (m, 2H), 2.94- 2.82 (m, 2H), 2.68-2.61 (m, 2H), 2.52(s, 3H), 2.17-1.91 (m, 4H), 1.82 (s, 3H), 1.76-1.71 (m, 2H), 1.65-1.59(m, 1H), 1.25- 1.21 (m, 3H), 0.55-0.51 (m, 2H), 0.49-0.46 (m, 2H). I-59

505.4 2.12 11.09 (CDCl₃) δ 8.51 (s, 1H), 7.59-7.58 (m, 1H), 7.40 (d, J =6.4 Hz, 1H), 7.16-7.11 (m, 2H), 6.52 (d, J = 6.8 Hz, 1H), 5.28-5.26 (m,1H), 5.04-5.02 (m, 1H), 3.71-3.57 (m, 3H), 3.38- 3.19 (m, 3H), 3.04-2.90(m, 4H), 2.53 (q, J = 7.2 Hz, 2H), 2.41-2.34 (m, 2H), 2.28-2.20 (m, 1H),2.14 (s, 3H), 2.05-2.01 (m, 2H), 1.95-1.92 (m, 2H), 1.79-1.64 (m, 2H),1.34- 1.22 (m, 9H), 1.16 (t, J = 7.2 Hz, 3H). I-60

491.4 2.83 10.49 (CD₃OD) δ 8.33 (d, J = 6.4 Hz, 1H), 7.63 (d, J = 8 Hz,1H), 7.38-7.32 (m, 2H), 7.24-7.21 (m, 1H), 6.79- 6.77 (m, 1H), 5.45 (d,J = 10.8 Hz, 1H), 5.16 (d, J = 10.8 Hz, 1H), 4.00 (t, J = 6.0 Hz, 1H),3.38-3.52 (m, 2H), 3.10-3.04 (m, 3H), 2.93-2.88 (m, 1H), 2.74- 2.48 (m,7H), 2.20-1.67 (m, 10H), 1.30-1.18 (m, 10H). I-61

449.4 2.16 7.85 (DMSO-d₆) δ 8.36 (s, 1H), 7.56 (d, J = 7.2 Hz, 1H), 7.28(d, J = 8.4 Hz, 1H), 7.20-7.16 (m, 2H), 6.61 (d, J = 6.4 Hz, 1H),5.40-5.36 (m, 1H), 5.07 (d, J = 13.2 Hz, 1H), 3.54 (br, 1H), 3.50-3.25(m, 4H), 3.22- 3.20 (m, 1H), 2.95-2.93 (m, 4H), 2.51-2.49 (m, 1H), 2.46(s, 3H), 2.42- 2.33 (m, 2H), 2.21-1.95 (m, 1H), 1.91-1.71 (m, 8H), 1.06(t, J = 7.2 Hz, 3H). I-62

434.4 1.33 8.27 (CDCl₃) δ 8.39-8.38 (m, 1H), 8.10-8.09 (m, 1H),7.57-7.53 (m, 1H), 7.41- 7.39 (m, 1H), 7.20-7.17 (m, 1H), 7.07-7.04 (m,1H), 6.82-6.79 (m, 1H), 5.05-5.02 (m, 2H), 4.80- 4.73 (m, 1H), 4.00-3.74(m, 2H), 2.98-2.82 (m, 3H), 2.58-2.35 (m, 6H), 2.18-1.50 (m, 14H). I-63

434.4 1.67 5.08 (CDCl₃) δ 8.40-8.37 (m, 2H), 7.62-7.52 (m, 2H),7.39-7.33 (m, 1H), 7.24- 7.20 (m, 1H), 7.07-6.97 (m, 1H), 5.50-5.00 (m,2H), 5.00-4.80 (m, 1H), 3.68-3.50 (m, 3H), 3.34- 3.32 (m, 1H), 3.00-2.50(m, 5H), 2.17 (s, 3H), 2.12-1.50 (m, 13H) I-66

431.4 1.94 9.61 (CDCl₃) δ 8.51 (d, J = 3.6 Hz, 1H), 7.63 (s, 1H), 7.43(dd, J = 7.6, 1.2 Hz, 1H), 7.31 (d, J = 8.8 Hz, 1H), 7.14-7.10 (m, 1H),7.05 (dd, J = 7.6, 4.8 Hz, 1H), 6.25 (dd, J = 7.2, 0.8 Hz, 1H),3.59-3.56 (m, 1H), 3.50-3.45 (m, 1H), 3.11 (s, 4H), 2.90 (s, 4H), 2.48(s, 3H), 2.09-2.05 (m, 2H), 1.97-1.93 (m, 2H), 1.91 (s, 3H), 1.83-1.78(m, 1H), 1.74-1.61 (m, 2H), 0.57- 0.52 (m, 2H), 0.51-0.46 (m, 2H). I-67

509.3 2.22 7.32 CDCl₃) δ 8.47 (s, 1H), 7.42-7.40 (m, 1H), 7.38 (dd, J =8.8, 0.8 Hz, 1H), 7.13-7.09 (m, 1H), 7.05 (dd, J = 7.6, 4.8 Hz, 1H),6.38 (dd, J = 7.2, 0.8 Hz, 1H), 3.63-3.60 (m, 2H), 3.30-3.24 (m, 2H),3.01- 2.98 (m, 2H), 2.88-2.76 (m, 4H), 2.47 (s, 3H), 2.23-1.90 (m, 4H),1.78 (s, 3H), 1.76-1.70 (m, 2H), 1.65-1.55 (m, 1H), 0.54- 0.50 (m, 2H),0.48-0.44 (m, 2H). I-68

431.4 2.28 8.44 (CD₃OD) δ 8.47 (d, J = 3.6 Hz, 1H), 7.61 (s, 1H), 7.42(dd, J = 7.2, 0.8 Hz, 1H), 7.31 (d, J = 9.2 Hz, 1H), 7.13-7.04 (m, 2H),6.28 (t, J = 3.6 Hz, 1H, ), 3.60 (d, J = 9.6 Hz, 1H, ), 3.48 (d, J = 7.6Hz, 1H), 3.29-3.17 (m, 4H), 3.07 (d, J = 10.4 Hz, 1H, ), 3.00 (d, J =10.8 Hz, 1H), 2.52 (s, 3H), 2.38 (s, 3H), 2.21-1.92 (m, 11H), 1.77-1.52(m, 2H) I-69

431.4 2.53 9.29 (CD₃OD) δ 8.49 (d, J = 3.6 Hz, 1H), 7.65 (s, 1H), 7.43(d, J = 7.2 Hz, 1H), 7.23 (d, J = 8.8 Hz, 1H), 7.10- 7.04 (m, 2H), 6.13(d, J = 6.8 Hz, 1H, ), 3.97 (s, 2H), 3.53-3.48 (m, 2H), 2.83 (d, J =10.4 Hz, 2H), 2.59 (d, J = 10.4 Hz, 2H), 2.50 (s, 3H), 2.38 (s, 3H),2.04- 1.87 (m, 11H), 1.77-1.64 (m, 2H) I-74

463.5 1.69 8.34 (CDCl₃) δ 8.54 (brs, 1H), 7.56 (d, J = 7.2 Hz, 1H), 7.33(d, J = 8.4 Hz, 1H), 7.14-7.07 (m, 2H, ), 6.41 (s, 1H), 5.81 (s, 1H),5.27 (d, J = 12.8 Hz, 1H), 3.78- 3.58 (m, 3H), 3.33 (s, 1H), 3.21 (d, J= 11.2 Hz, 1H), 3.06 (t, J = 11.2 Hz, 1H), 2.93 (d, J = 11.6 Hz, 2H),2.75 (s, 1H), 2.61-2.56 (m, 1H), 2.51-2.44 (m, 1H), 2.42 (s, 3H), 2.35(s, 1H), 2.17-2.14 (m, 1H), 2.05- 1.93 (m, 3H), 1.87 (s. 3H), 1.78 (d, J= 11.6 Hz, 1H), 1.67-1.58 (m, 1H), 1.26 (d, J = 6.8 Hz, 6H). I-75

477.4 1.77 4.69 (CDCl₃) δ 8.46-8.44 (s, 1H), 7.52 (s, 1H), 7.28- 7.26(m, 1H), 7.07-7.05 (m, 2H), 6.40-6.36 (m, 1H), 5.75 (brs, 1H), 5.23-5.21 (m, 1H), 3.56-3.23 (m, 3H), 3.24-3.19 (m, 2H), 3.10-2.96 (m, 3H),2.79-2.72 (m, 1H), 2.60- 2.37 (m, 4H), 2.22-2.08 (m, 1H), 2.00-1.94 (m,3H), 1.87-1.82 (m, 4H), 1.63-1.47 (m, 2H), 1.20 (d, J = 7.2 Hz, 6H),1.12 (t, J = 7.2 Hz, 3H). I-76

420.4 1.66 8.05 (CDCl₃) δ 8.50 (d, J = 3.6 Hz, 1H), 7.55 (1H, s), 7.62(dd, J = 7.6, 1.2 Hz, 1H), 7.32 (dd, J = 7.6, 1.2 Hz, 1H), 6.31 (d, J =7.6 Hz, 1H), 6.15 (d, J = 7.6 Hz, 1H), 4.30 (brs, 2H), 3.57 (dd, J =10.8, 1.6 Hz, 1H, ), 3.42 (dd, J = 10.8, 1.6 Hz, 1H), 3.05 (s, 4H), 2.66(s, 4H), 2.44-2.41 m, 6H), 2.03-1.87 m, 4H), 1.73- 1.59 (m, 5H). I-77

431.4 1.81 6.33 (CDCl₃) δ 8.51 (d, J = 3.6 Hz, 1H), 7.63 (s, 1H), 7.43(dd, J = 7.6, 1.2 Hz, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.13 (dd, J = 8.8,7.2 Hz, 1H), 7.05 (dd, J = 7.6, 4.8 Hz, 1H), 6.30 (d, J = 7.2 Hz, 1H),3.59-3.52 (m, 2H), 3.49-3.42 (m, 2H), 3.21-3.17 (m, 2H), 3.00- 2.89 (m,1H), 2.68-2.55 (m, 2H), 2.48 (s, 3H), 2.41-2.29 (m, 2H), 2.09- 2.03 (m,3H), 1.98-1.93 (m, 4H), 1.87-1.71 (m, 4H), 1.68-1.47 (m, 2H). I-78

433.3 1.60 6.04 (CDCl₃) δ 8.50 (d, J = 4 Hz, 1H), 7.57 (s, 1H), 7.42 (d,J = 6.8 Hz, 1H), 7.31 (d, J = 8.8 Hz, 1H), 7.12 (dd, J = 8.8, 7.2 Hz,1H), 7.05 (dd, J = 7.6, 4.8 Hz, 1H), 6.24 (d, J = 6.4 Hz, 1H), 3.58-3.54(m, 2H), 3.52-3.45 (m, 2H), 2.76- 2.68 (m, 2H), 2.48 (s, 3H), 2.45-2.37(m, 1H), 2.39 (s, 6H), 2.08-1.92 (m, 4H), 1.90 (s, 3H), 1.88-1.85 (m,2H), 1.77-1.61 (m, 4H). I-79

431.4 1.76 8.82 (CDCl₃) δ 8.49 (s, 1H), 7.51 (s, 1H), 7.33-7.27 (m, 2H),7.14 (dd, J = 9.0, 7.0 Hz, 1H), 7.06 (dd, J = 8.0, 4.8 Hz, 1H), 6.27(dd, J = 7.2, 0.8, 1H), 3.97 (d, J = 8.4 Hz, 1H), 3.48 (dd, J = 11.6,2.4 Hz, 1H), 3.18- 3.14 (m, 4H), 2.77-2.70 (m, 4H), 2.43 (s, 3H),2.13-2.05 (m, 3H), 1.99- 1.94 (2H, m), 1.89 (s, 3H), 1.77 (d, J = 9.2Hz, 1H), 1.67-1.62 (m, 2H), 1.07- 1.00 (m, 2H), 0.72-0.69 (m, 1H),0.68-0.51 (m, 1H). I-80

459.3 1.83 9.16 (CDCl₃) δ 8.90 (d, J = 3.6 Hz, 1H), 7.95 (dd, J = 7.6,1.2 Hz, 1H), 7.61 (s, 1H), 7.34-7.27 (m, 2H), 7.14 (dd, J = 12.0, 8.0Hz, 1H), 6.28 (d J = 6.8 Hz, 1H), 3.73 (d, J = 11.2 Hz, 1H), 3.52 (dd, J= 11.6, 2.4 Hz, 1H), 3.15 (s, 4H), 2.70 (s, 4H), 2.43 (s, 4H), 2.18-1.94 (m, 4H), 1.86 (s, 3H), 1.81-1.78 (m, 1H). I-81

475.3 1.84 9.14 (CDCl₃) δ 8.65 (dd, J = 4.4, 1.2 Hz, 1H), 7.59 (s, 1H),7.58-7.56 (m, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.25-7.23 (m, 1H),7.16-7.12 (m, 1H), 6.28 (dd, J = 7.2, 0.8 Hz, 1H), 3.77 (dd, J = 11.2,2.4 Hz, 1H), 3.49 (dd, J = 11.2, 2.8 Hz, 1H), 3.15 (s, 4H), 2.69 (s,4H), 2.43 (s, 3H), 2.07-1.93 (m, 3H), 1.88 (s, 3H), 1.73-1.62 (m, 3H).I-82

425.3 1.78 8.85 (CDCl₃) δ 8.61 (dd, J = 4.4, 1.2 Hz, 1H), 7.67 (dd, J =8.0, 1.6 Hz, 1H), 7.62 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.16-7.11 (m,2H), 6.28 (d, J = 7.2 Hz, 1H), 3.97 (dd, J = 10.8, 2.8 Hz, 1H), 3.53(dd, J = 11.2, 2.8 Hz, 1H), 3.16 (brs, 4H), 2.70 (s, 4H), 2.43 (s, 3H),2.10-1.94 (m, 4H), 1.92 (s, 3H), 1.72- 1.65 (m, 2H). I-83

416.4 1.69 8.44 (CDCl₃) δ 8.44 (s, 1H), 7.78 (d, J = 6.8 Hz, 1H), 7.60(d, J = 8.8 Hz, 1H), 7.41 (d, J = 7.2 Hz, 1H), 7.14 (t, J = 7.6 Hz, 1H),7.05 (dd, J = 7.6, 4.8 Hz, 1H), 6.77 (t, J = 6.8 Hz, 1H), 6.19 (s, 1H),3.65- 3.49 (m, 1H), 3.48-3.34 (m, 1H), 2.71-2.39 (m, 8H), 2.34 (s, 3H),2.34- 2.25 (m, 1H), 2.16-1.90 (m, 4H), 1.81-1.68 (6H, m), 1.54-1.60 (m,1H). I-85

475.3 2.09 6.92 (CDCl₃) δ 8.47 (s, 1H), 7.41 (d, J = 8.8 Hz, 1H), 7.20(dd, J = 9.2, 7.6 Hz, 1H), 7.05 (dd, J = 7.6, 4.8 Hz, 1H), 6.51 (d, J =6.8 Hz, 1H), 5.35-5.30 (m, 1H), 5.12 (d, J = 8.8 Hz, 1H), 3.57-3.50 (m,2H), 3.41 (s, 1H), 3.27-3.20 (m, 2H), 3.01-2.96 (m, 1H), 2.93-2.88 (m,3H), 2.49- 2.43 (m, 4H), 2.17-1.99 (m, 4H), 1.78 (s, 6H), 1.62-1.55 (m,1H), 1.28- 0.88 (m, 2H), 0.65-0.61 (m, 2H), 0.53-0.48 (m, 2H). I-91

467.3 2.09 10.84 (CDCl₃) δ 8.48 (brs, 1H), 7.58 (d, J = 3.6 Hz, 1H),7.33 (d, J = 8.8 Hz, 1H), 7.14-7.08 (m, 2H), 6.35 (d, J = 7.2 Hz, 1H),3.63 (d, J = 50.0 Hz, 2H), 3.30 (t, J = 22.4 Hz, 2H), 2.94 (s, 5H),2.87-2.84 (m, 3H), 3.55- 3.47(m, 2H), 2.40 (s, 3H), 2.28-2.17 (m, 1H),2.06 (brs, 1H), 1.96-1.93 (m, 1H), 1.78 (s, 2H), 1.67- 1.56 (m, 1H),1.27-1.23 (m, 6H). I-88

453.3 2.03 7.29 (CDCl₃) δ 8.46 (s, 1H), 7.52-7.32 (m, 2H), 7.12- 7.03(m, 2H), 6.35 (d, J = 7.2 Hz, 1H), 3.61-3.52 (m, 2H), 3.35-3.29 (m, 2H),2.98-2.87 (m, 7H), 2.24- 2.06 (m, 3H), 1.82-1.77 (m, 4H), 1.76-1.61 (m,2H), 1.16 (t, J = 7.0 Hz, 3H). I-92

467.4 2.15 7.57 (CDCl₃) δ 8.42 (s, 1H), 7.41 (d, J = 6.8 Hz, 1H), 7.33(d, J = 8.8 Hz, 1H), 7.12-7.03 (m, 2H), 6.35- 6.33 (m, 1H), 3.61-3.53(m, 2H), 3.36-3.30 (m, 2H), 2.93-2.83 (m, 4H), 2.78-2.72 (m, 1H), 2.68-2.52 (m, 5H), 2.27-2.17 (m, 1H), 2.07-1.92 (m, 2H), 1.91-1.79 (m, 4H),1.75-1.31 (m, 2H), 1.12 (d, J = 6.4 Hz, 6H). I-93

465.3 2.22 7.34 (CDCl₃) δ 8.47 (s, 1H), 7.41 (dd, J = 7.6, 0.8 Hz, 1H),7.34 (dd, J = 8.8, 0.8 Hz, 1H), 7.11-7.03 (m, 2H), 6.32 (dd, J = 7.2,0.8 Hz, 1H), 3.62-3.55 (m, 2H), 3.32-3.25 (m, 2H), 3.03-3.00 (m, 2H),2.89- 2.69 (m, 4H), 2.47 (s, 3H), 2.25-1.92 (m, 4H), 1.79 (s, 3H),1.77-1.71 (m, 2H), 1.64-1.57 (m, 1H), 0.54- 0.50 (m, 2H), 0.48-0.44 (m,2H). I-89

429.4 1.76 5.60 (CDCl₃) δ 8.47 (s, 1H), 7.41-7.38 (m, 2H), 7.21- 7.17(m, 1H), 7.06-7.03 (m, 1H), 6.40 (d, J = 7.2 Hz, 1H), 3.83 (s, 1H),3.70-3.58 (m, 2H), 3.47- 3.38 (m, 2H), 2.98-2.84 (m, 2H), 2.83-2.79 (m,2H), 2.63-2.54 (m, 2H), 2.52-2.40 (m, 3H), 2.38 (s, 3H), 2.10-2.03 (m,2H), 1.80 (s, 3H), 1.73-1.61 (m, 2H), 1.58-1.24 (m, 2H). I-104

448.4 0.59 4.41 (CD₃OD) δ 8.45-8.27 (m, 2H), 7.54-7.46 (m, 2H),7.26-7.23 (m, 1H), 7.15- 7.12 (m, 1H), 6.85-6.82 (m, 1H), 4.60-4.30 (m,1H), 3.03 (s, 3H), 3.00- 2.40 (m, 4H), 2.24 (s, 3H), 2.20-1.20 (m, 19H).I-105

448.4 0.57 4.31 (CD₃OD) δ 8.48 (d, J = 6.8 Hz, 1H), 8.23 (br, 1H),7.53-7.48 (m, 2H), 7.27- 7.12 (m, 2H), 6.85-6.81 (m, 1H), 4.60-4.50 (m,1H), 3.39-3.38 (m, 1H), 3.03 (s, 3H), 3.00-2.50 (m, 4H), 2.25-2.22 (m,5H), 2.21 (s, 3H), 2.00-1.20 (m, 13H). I-106

423.4 1.73 8.43 (CDCl₃) δ 8.51 (d, J = 4.0 Hz, 1H), 7.71 (s, 1H), 7.44-7.42 (m, 1H), 7.32-7.29 (m, 1H), 7.07-7.02 (m, 2H), 4.90-4.30 (m, 2H),3.58-3.54 (m 1H) 3.45- 3.42 (m, 1H), 3.40-3.25 (m, 4H), 2.92-2.79 (m,2H), 2.46 (s, 3H), 2.43 (s, 3H), 2.04-1.93 (m, 4H), 1.87 (s, 3H),1.64-1.60 (m, 2H). I-109

417.4 2.01 4.89 (CD₃OD) δ 8.44 (s, 1H), 7.98 (s, 1H), 7.61 (d, J = 7.2Hz, 1H), 7.32-7.18 (m, 3H), 6.51 (dd, J = 6.8, 0.8 Hz, 1H), 3.66-3.63(m, 3H), 3.48-3.44 (m, 1H), 3.32-3.29 (m, 2H), 3.04- 2.97 (m, 2H), 2.53(s, 3H), 2.32-2.27 (m, 2H), 2.02- 1.88 (m, 6H), 1.78 (s, 3H), 1.76-1.68(m, 2H). I-110

417.4 2.00 5.61 (CDCl₃) δ 8.41 (d, J = 4.0 Hz, 1H), 7.58 (s, 1H),7.36-7.34 (m, 1H), 7.18- 7.16 (m, 1H), 7.03-6.97 (m, 2H), 6.05 (d, J =6.8 Hz, 1H), 3.85 (d, J = 3.2 Hz, 2H), 3.53 (dd, J = 11.2, 2.8 Hz, 1H),3.42 (dd, J = 10.8, 2.8 Hz, 1H), 3..27 (t, J = 11.6 Hz, 2H), 2.814-2.78(m, 2H), 2.42 (s, 3H), 2.03-1.85 (m, 11H), 1.71-1.57 (m, 2H). I-111

447.4 1.96 6.13 (CDCl₃) δ 8.40 (s, 1H), 7.53 (d, J = 7.6 Hz, 2H), 7.25(d, J = 9.2 Hz 1H), 7.09-7.05 (m, 2H), 6.21 (d, J = 6.4 Hz, 1H),3.62-3.60 (m, 1H), 3.58-3.56 (m, 1H), 3.12-3.10 (m, 4H), 2.66 (m, 3H),2.48 (q, J = 7.2 Hz, 2H), 1.95 (m, 1H), 1.89-1.86 (m, 4H), 1.78 (s, 3H),1.71-1.53 (m, 2H), 1.26-1.19 (m, 6H), 1.10 (t, J = 7.6 Hz, 3H). I-112

461.4 1.73 8.55 (CDCl₃) δ 8.54 (s, 1H), 7.41 (d, J = 7.2 Hz, 1H), 7.33(d, J = 8.4 Hz, 1H), 7.11-7.03 (m, 2H), 6.40 (d, J = 2.4 Hz, 1H), 5.82(s, 1H), 5.29 (d, J = 14.0 Hz, 1H), 3.67-3.61 (m, 2H), 3.55 (s, 1H),3.21 (d, J = 11.2 Hz, 1H), 3.09 (d, J = 11.6 Hz, 2H), 3.01-2.94 (m, 1H),2.83-2.67 (m, 3H), 2.40 (s, 3H), 2.17- 2.12 (m, 1H), 2.05-1.94 (m, 3H),1.88 (s, 3H), 1.80-1.78 (m, 3H), 1.66- 1.56 (m, 1H), 0.55-0.50 (m, 2H),0.49-0.44 (m, 2H). I-113

465.3 2.10 10.80 (CDCl₃) δ 8.47 (d, J = 2.0 Hz, 1H), 7.41 (dd, J = 7.6,0.8 Hz, 1H), 7.36-7.33 (m, 1H), 7.12-7.03 (m, 2H), 6.38 (d, J = 7.2 Hz,1H), 3.61-3.55 (m, 2H), 3.48- 3.42 (m, 1H), 3.38-3.31 (m, 1H), 3.18-3.08(m, 2H), 2.97-2.83 (m, 1H), 2.68-2.55 (m, 2H), 2.46 (s, 3H), 2.44-2.41(m, 1H), 2.38-2.30 (m, 1H), 2.27- 2.14 (m, 2H), 2.07-1.82 (m, 4H), 1.78(s, 3H), 1.74-1.64 (m, 2H), 1.61- 1.42 (m, 2H) I-114

467.3 1.81 4.97 (CDCl₃) δ 8.38 (s, 1H), 7.34 (d, J = 7.6 Hz, 1H), 7.26(d, J = 8.4 Hz, 1H), 7.04-6.96 (m, 2H), 6.25- 6.23 (m, 1H), 3.60-3.51(m, 5H), 3.41-3.36 (m, 2H), 2.65-2.55 (m, 2H), 2.43 (m, 2H), 2.36 (s,9H), 2.14-2.10 (m, 1H), 1.97- 1.93 (m, 2H), 1.90-1.80 (m, 2H), 1.74 (s,3H), 1.56-1.52 (m, 1H). I-115

459.3 2.03 10.38 (CDCl₃) δ 8.59 (dd, J = 4.4, 1.2 Hz, 1H), 7.65 (dd, J =8.0, 1.2 Hz, 1H), 7.34 (dd, J = 8.8, 0.8 Hz, 1H), 7.13-7.08 (m, 2H),6.35 (dd, J = 7.2, 0.8 Hz, 1H), 3.98 (d, J = 9.2 Hz, 1H), 3.62 (dd, J =14.4, 2.8 Hz, 1H), 3.33-3.26 (m, 2H), 2.98-2.84 (m, 4H), 2.56- 2.46 (m,2H), 2.41 (s, 3H), 2.02-1.93 (m, 2H), 1.82 (s, 3H), 1.78-1.75 (m, 2H),1.68-1.57 (m, 1H), 1.26 (s, 1H). I-116

455.4 1.63 7.87 (CDCl₃) δ 8.61 (d, J = 4.0 Hz, 1H), 7.65 (dd, J = 8.0,Hz, 1H), 7.33 (d, J = 8.8 Hz, 1H), 7.14-7.08 (m, 2H), 6.42 (d, J = 7.2Hz, 1H), 5.80 (s, 1H), 5.25 (d, J = 14.8 Hz, 1H), 4.00 (d, J = 10.0 Hz,1H), 3.70 (dd, J = 11.2, Hz, 1H), 3.56 (s, 1H), 3.20 (d, J = 9.2 Hz,1H), 3.10-3.03 (m, 1H), 2.93 (d, J = 12.0 Hz, 2H), 2.77 (t, J = 8.4 Hz,1H), 2.61-2.54 (m, 1H), 2.50- 2.43 (m, 1H), 2.41 (3H, s), 2.19-1.93 (m,4H), 1.91 (s, 3H), 1.82-1.80 (m, 2H). I-117

483.3 1.95 9.86 (CDCl₃) δ 8.58 (d, J = 4.0 Hz, 1H), 7.65 (dd, J = 8.0Hz, J = 1.2 Hz, 1H), 7.40 (d, J = 8.0 Hz, 1H), 7.14- 7.10 (m, 2H), 6.50(d, J = 6.8 Hz, 1H), 5.30-5.28 (m, 1H), 5.08 (s, 1H), 3.94 (s, 1H), 3.66(q, J = 7.2 Hz, 2H), 3.58 (d, J = 12.0 Hz, 1H), 3.26 (t, J = 12.0 Hz,2H), 3.02-2.87 (m, 4H), 2.40 (s, 3H), 2.19-2.15 (m, 1H), 2..03 (s, 6H),1.82 (s, 3H), 1.66-1.58(m, 1H), 1.22 (t, J = 6.8 Hz, 3H). I-119

493.3 2.05 10.51 (CDCl₃) δ 8.88 (d, J = 3.2 Hz, 1H), 7.93 (d, J = 6.8Hz, 1H), 7.35-7.32 (m, 2H), 7.12-7.07 (m, 1H), 6.35 (d, J = 6.4 Hz, 1H),3.75 (d, J = 10.4 Hz, 1H), 3.64-3.59 (m, 1H), 3.32- 3.25 (m, 2H),2.93-2.83 (m, 4H), 2.52-2.49 (m, 2H), 2.40 (s, 3H), 2.23- 2.22 (m, 1H),1.96-1.78 (m, 2H), 1.76 (s, 3H), 1.64-1.60 (m, 1H). I-123

487.2 2.59 7.67 (CDCl₃): δ 8.60-8.58 (m, 1H), 7.66-7.64 (m, 1H), 7.35(d, J = 8.8 Hz, 1H), 7.14-7.09 (m, 2H), 6.39 (d, J = 7.2 Hz, 1H),4.00-3.97 (m, 1H), 3.64-3.60 (m, 1H), 3.02-2.73 (m, 9H), 1.99-1.94 (m,3H), 1.81- 1.78 (m, 6H), 1.25 (brs, 6H). I-125

473.2 2.45 11.75 (CDCl₃): δ 8.51-8.50 (m, 1H), 7.58-7.56 (m, 1H),7.27-7.25 (m, 1H), 7.05- 7.00 (m, 2H), 6.28 (d, J = 6.8 Hz, 1H),3.91-3.89 (m, 1H), 3.56-3.52 (m, 1H), 3.28-3.20 (m, 2H), 2.90- 2.80 (m,4H), 2.50-2.36 (m, 4H), 2.23 (brs, 2H), 1.91-1.85 (m, 2H), 1.73 (s, 3H),1.73-1.66 (m, 2H), 1.10-1.07 (m, 3H). I-126

433.4 2.27 8.52 (CDCl₃) δ 8.34 (d, J = 3.6 Hz, 1H), 7.65 (s, 1H),7.52-7.48 (m, 1H), 7.24- 7.10 (m, 3H), 6.41 (d, J = 6.0 Hz, 1H), 4.08(dd, J = 11.2 Hz, 1H), 3.86 (t, J = 6.4 Hz, 1H), 3.07 (br, 4H),2.78-2.73 (m, 1H), 2.67- 2.54 (m, 7H), 2.33 (s, 3H), 1.95-1.33 (m, 6H),0.66 (t, J = 7.2 Hz, 6H). I-127

467.3 2.78 10.37 (CDCl₃) δ 8.38 (d, J = 3.6 Hz, 1H), 7.34-7.28 (m, 2H),7.19-6.96 (m, 2H), 6.29 (t, J = 7.6 Hz, 1H), 3.99 (br, 2H), 3.24-3.20(m, 2H), 2.88-2.78 (m, 4H), 2.64-2.45 (m, 6H), 2.33 (s, 3H), 2.09-1.65(m, 6H), 0.68 (dd, J = 6.8 Hz, 6H). I-132

487.2 1.90 5.79 (CDCl₃) δ 8.59-8.58 (m, 1H), 7.66-7.64 (m, 1H), 7.32 (d,J = 8.8 Hz, 1H), 7.14-7.07 (m, 2H), 6.31 (d, J = 6.8 Hz, 1H), 4.00-3.97(m, 1H), 3.63 (dd, J = 11.6, 2.8 Hz, 1H), 3.46- 3.40 (m, 2H), 2.72-2.59(m, 2H), 2.35 (s, 6H), 2.30-2.23 (m, 1H), 2.22- 2.10 (m, 1H), 1.98-1.96(m, 1H), 1.90-1.82 (s, 4H), 1.80 (s, 3H), 1.78-1.74 (m, 2H), 1.68-1.60(m, 2H). I-134

481.3 2.40 8.75 (CD₃OD): δ 8.60 (d, J = 4.0 Hz, 1H), 7.86-7.84 (m, 1H),7.32-7.29 (m, 1H), 7.22-7.21 (m, 2H), 6.57- 6.56 (m, 1H), 6.00-5.95 (m,1H), 5.31-5.27 (m, 1H), 4.07-4.00 (m, 3H), 3.66 (d, J = 11.2 Hz, 1H),2.91-2.84 (m, 2H), 2.77 (d, J = 11.6 Hz, 1H), 2.63 (d, J = 10.4 Hz, 1H),2.39 (s, 3H), 2.17 (s, 3H), 2.15- 2.10 (m, 2H), 2.03-1.99 (m, 2H),1.96-1.94 (m, 4H), 1.91-1.69 (m, 2H). I-135

481.4 2.19 7.89 (DMSO-d₆): δ 8.58 (dd, J = 4.4, 1.2 Hz, 1H), 7.93-7.91(m, 1H), 7.34 (dd, J = 8.0, 4.8 Hz, 1H), 7.27 (d, J = 7.6 Hz, 1H), 7.15(dd, J = 8.8, 7.2 Hz, 1H), 6.76 (d, J = 6.4 Hz, 1H), 5.63-5.59 (m, 1H),5.14 (d, J = 9.2 Hz, 1H), 3.93-3.90 (m, 1H), 3.59-3.56 (m, 1H),3.22-3.13 (m, 4H), 2.99- 2.97 (d, J = 7.6 Hz, 1H), 2.78-2.76 (m, 1H),2.25 (s, 3H), 2.09-1.97 (m, 4H), 1.90-1.80 (m, 4H), 1.71 (s, 3H),1.60-1.57 (m, 2H). I-136

483.3 1.56 7.67 (CDCl₃) δ 8.62 (d, J = 4.0 Hz, 1H), 7.66-7.64 (m, 1H),7.28 (d, J = 8.4 Hz, 1H), 7.14-7.07 (m, 2H), 6.40 (d, J = 6.8 Hz, 1H),5.72 (s, 1H), 5.30-5.21 (m, 1H), 4.00 (s, 1H), 3.70- 3.62 (m, 2H),3.38-3.35 (m, 1H), 2.81 (t, 1H), 2.60- 2.57 (m, 1H), 2.45-2.40 (m, 1H),2.36 (s, 6H), 2.29-2.22 (m, 1H), 2.20- 2.10 (m, 1H), 2.01-1.94 (m, 4H),1.91 (s, 3H), 1.89-1.80 (m, 3H), 1.68- 1.54 (m, 2H). I-137

465.4 1.94 9.78 (CDCl₃) δ 8.47 (s, 1H), 7.42-7.36 (m, 2H), 7.11- 7.03(m, 2H), 6.64 (d, J = 6.8 Hz, 1H), 3.61-3.55 (m, 2H), 3.23-3.05 (m, 6H),2.47 (s, 3H), 2.36 (s, 3H), 2.29-2.16 (m, 4H), 2.06- 1.90 (m, 4H), 1.78(s, 3H), 1.74-1.64 (m, 1H), 1.61- 1.56 (m, 1H). I-138

465.4 2.12 10.81 (CDCl₃) δ 8.45 (s, 1H), 7.41 (dd, J = 7.4, 1.0 Hz, 1H),7.24 (s, 3H), 7.07- 7.00 (m, 2H), 6.23 (d, J = 7.2 Hz, 1H), 3.86 (s,2H), 3.58 (t, J = 12.6 Hz, 2H), 2.74-2.69 (m, 4H), 2.52 (brs, 3H), 2.32(s, 3H), 2.23-2.19 (m, 1H), 2.06- 1.92 (m, 6H), 1.80 (s, 3H), 1.75-1.71(m, 2H), 1.62- 1.58 (m, 1H). I-139

463.4 1.61 8.03 (CDCl₃) δ 8.59 (brs, 1H), 7.42 (d, J = 7.2 Hz, 1H), 7.35(d, J = 6.4 Hz, 1H), 7.13-7.10 (m, 1H), 7.09- 7.04 (m, 1H), 6.51 (brs,1H), 5.65 (d, J = 4.4 Hz, 1H), 3.74-3.72 (m, 1H), 3.62-3.58 (m, 1H),3.29- 3.27 (m, 1H), 3.18-3.09 (m, 2H), 2.94-2.91 (m, 1H), 2.79 (d, J =8.8 Hz, 1H), 2.62-2.40 (m, 3H), 2.38 (s, 3H), 2.32 (s, 3H), 2.11- 1.92(m, 4H), 1.92 (s, 3H), 1.70-1.60 (m, 4H), 1.06 (t, J = 7.2 Hz, 3H).I-140

464.3 1.66 5.65 (CDCl₃) δ 8.55-8.10 (m, 1H), 7.40 (d, J = 6.4 Hz, 1H),7.35 (d, J = 8.8 Hz, 1H), 7.10-7.03 (m, 2H), 6.50 (d, J = 6.8 Hz, 1H),5.32 (s, 1H), 3.70 (d, J = 3.2 Hz, 1H), 3.59 (d, J = 8.0 Hz, 1H),3.32-3.18 (m, 2H), 3.12-3.08 (m, 1H), 2.93 (d, J = 10.4 Hz, 1H), 2.80(d, J = 9.6 Hz, 1H), 2.63-2.42, (m, 4H), 2.37 (s, 3H), 2.00 (d, J = 12.8Hz, 2H), 1.93 (s, 3H), 1.90-1.64 (4H, m), 1.33- 1.27 (m, 1H), 1.22 (s,3H). I-141

445.4 1.95 6.39 (CDCl₃) δ 8.47 (d, J = 2.8 Hz, 1H), 7.39 (dd, J = 7.4,1.0, 1H), 7.33 (dd, J = 8.8, 0.8 Hz, 1H), 7.09 (dd, J = 15.6, 1.6 Hz,1H), 7.06- 7.01 (m, 2H), 6.28 (dd, J = 7.2, 0.8 Hz, 1H), 6.79-5.72 (m,1H), 3.56 (dd, J = 11.6, 2.8 Hz, 2H), 3.29 (dd, J = 1.08, 2.2 Hz, 1H),3.17 (dd, J = 11.0, 2.6 Hz, 1H), 3.01-2.94 (m, 1H), 2.86 (dd, J = 11.2,2.0 Hz, 1H), 2.79 (dd, J = 11.2, 1.6 Hz, 1H), 2.74-2.67 (m, 1H), 2.48(brs, 2H), 2.37 (s, 3H), 2.36-2.24 (m, 3H), 2.07-1.93 (m, 4H), 1.86-1.75 (m, 2H), 1.71 (s, 3H), 1.63-1.55 (m, 3H). I-142

453.3 1.66 8.49 (CDCl₃) δ 8.61-8.60 (m, 1H), 7.66 (dd, J = 8.0, 1.6 Hz,1H), 7.58 (s, 1H), 7.31 (d, J = 8.8 Hz, 1H), 7.14- 7.11 (m, 2H),6.25-6.23 (m, 1H), 3.98-3.95 (m, 1H), 3.54-3.49 (m, 3H), 2.76-2.68 (m,2H), 2.45- 2.41 (m, 1H), 2.39 (s, 6H), 2.12-2.04 (m, 1H), 2.05- 1.96 (m,4H), 1.92 (s, 3H), 1.86-1.81 (m, 3H), 1.78- 1.63 (m, 2H). I-146

425.4 1.70 8.21 (CDCl₃) δ 8.40 (dd, J = 4.8, 1.2 Hz, 1H), 7.38 (dd, J =7.2, 1.6 Hz, 1H), 7.32 (dd, J = 9.2, 1.2 Hz, 1H), 7.08 (dd, J = 8.8, 7.2Hz, 1H), 7.02 (dd, J = 8.0, 4.8 Hz, 1H), 6.34 (dd, J = 7.2, 0.8 Hz, 1H),4.25-4.18 (m, 2H), 3.29-3.25 (m, 2H), 2.96-2.78 (m, 4H), 2.53-2.46 (m,2H), 2.38 (s, 6H), 2.16-2.11 (m, 1H), 1.88-1.80 (m, 4H), 1.72- 1.65 (m,2H). I-147

445.2 3.22 9.15 (CDCl₃) δ 8.48 (dd, J = 4.0, 1.6, 1H), 7.63-7.60 (m,1H), 7.33 (dd, J = 8.8, 1.2 Hz, 1H), 7.11-7.07 (m, 2H), 6.35 (dd, J =7.2, 0.8 Hz, 1H), 4.52-4.48 (m, 1H), 4.25 (dd, J = 10.0, 3.6 Hz, 1H),3.31-3.26 (m, 2H), 2.97-2.91 (m, 1H), 2.86-2.79 (m, 3H), 2.54- 2.46 (m,2H), 2.39 (s, 3H), 2.15-2.12 (m, 1H), 2.02- 1.98 (m, 1H), 1.93-1.85 (m,3H), 1.54-1.44 (m, 1H). I-188

411.2 1.60 5.63 (CDCl₃) δ 8.59-8.50 (dd, J = 4.8, 1.6 Hz, 1H), 7.65-7.63 (m, 1H), 7.51 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.17-7.10 (m, 2H),6.27 (dd, J = 7.2, 0.8 Hz, 1H), 4.52 (dd, J = 11.2, 2.4, 1H), 4.20-4.17(m, 1H), 3.14 (s, 4H), 2.66 (m, 4H), 2.40 (s, 3H), 2.23-2.19 (m, 1H),2.15-2.11 (m, 1H), 2.02-1.98 (m, 1H), 1.89- 1.77 (m, 2H), 1.61-1.54 (m,1H). I-189

411.2 1.60 5.63 (CDCl₃) δ 8.59-8.50 (dd, J = 4.8, 1.6 Hz, 1H), 7.65-7.63 (m, 1H), 7.51 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.17-7.10 (m, 2H),6.27 (dd, J = 7.2, 0.8 Hz, 1H), 4.52 (dd, J = 11.2, 2.4, 1H), 4.20-4.17(m, 1H), 3.14 (s, 4H), 2.66 (m, 4H), 2.40 (s, 3H), 2.23-2.19 (m, 1H),2.15-2.11 (m, 1H), 2.02-1.98 (m, 1H), 1.89- 1.77 (m, 2H), 1.61-1.54 (m,1H). I-150

453.2 2.07 6.39 (CD₃OD): δ 8.51-8.50 (m, 1H), 7.93 (s, 1H), 7.77- 7.74(m, 1H), 7.23-7.13 (m, 3H), 6.38-6.36 (m, 1H), 3.92-3.88 (m, 1H),3.40-3.36 (m, 1H), 3.11 (brs, 4H), 2.80-2.70 (m, 5H), 1.94-1.54 (m, 9H),1.11-1.04 (m, 6H). I-151

439.2 1.95 6.22 (CD₃OD): δ 8.62 (dd, J = 4.4, 1.6 Hz, 1H), 8.02 (s, 1H),7.88 (dd, J = 8.2, 1.2 Hz, 2H), 7.35-7.25 (m, 3H), 6.51 (dd, J = 7.2,1.0 Hz, 1H), 4.04-3.99 (m, 1H), 3.51-3.49 (m, 1H), 3.24 (brs, 4H), 2.84(brs, 4H), 2.67-2.59 (m, 2H), 2.06-1.67 (m, 9H), 1.66- 1.23 (m, 3H).I-154

391.3 1.53 7.07 (CDCl₃) δ 8.40 (dd, J = 4.4, 0.8 Hz, 1H), 8.04 (d, J =6.8 Hz, 1H), 7.49 (d, J = 9.2 Hz, 1H), 7.37 (dd, J = 7.6, 0.8 Hz, 1H),7.09-7.05 (m, 1H), 7.00 (dd, J = 8.0, 4.8 Hz, 1H), 6.73-6.69 (m, 1H),4.29-4.23 (m, 1H), 4.16-4.11 (m, 1H), 3.43- 3.11 (m, 4H), 3.10-2.72 (m,4H), 2.41 (s, 3H), 2.36 (s, 3H), 2.14-2.10 (m, 1H), 1.90-1.76 (m, 4H),1.67- 1.59 (m, 2H). I-164

454.4 1.89 9.55 (CDCl₃) δ 8.48 (d, J = 4.0 Hz, 1H), 7.41 (dd, J = 8.0,1.6 Hz, 1H), 7.04 (dd, J = 8.0, 1.6 Hz, 1H), 6.30-6.24 (m, 2H), 4.42 (s,2H), 3.63-3.55 (m, 2H), 3.45- 3.31 (m, 2H, ), 3.21-3.14 (m, 2H),3.16-2.80 (m, 3H), 2.70-2.61 (m, 3H, ), 2.54-2.42 (m, 2H), 2.10- 1.94(m, 2H), 1.92-1.85 (m, 3H), 1.72-1.65 (m, 2H). I-166

423.2 1.58 7.40 (CDCl₃) δ 8.50 (dd, J = 4.8, 1.6 Hz, 1H), 7.65 (dd, J =8.0, 1.6 Hz, 1H), 7.57 (s, 1H), 7.33 (d, J = 8.8 Hz, 1H), 7.16-7.10 (m,2H), 6.24 (dd, J = 7.2, 0.8 Hz, 1H), 4.54 (dd, J = 11.2, 2.4 Hz, 1H),4.20 (dd, J = 11.2, 2.4 Hz, 1H), 3.18 (t, J = 4.8 Hz, 2H), 3.08 (s, 2H),2.93 (s, 2H), 2.23 (dd, J = 12.4, 2.4 Hz, 1H), 2.16-2.11 (m, 1H), 2.03(dd, J = 12.8, 2.0 Hz, 1H), 1.92-1.78 (m, 4H), 1.59-1.49 (m, 1H), 0.76(t, J = 6.0 Hz, 2H), 0.66 (t, J = 4.4 Hz, 2H). I-173

441.3 1.68 8.09 (CDCl₃) δ 8.16 (dd, J = 4.0, 2.0 Hz, 1H), 7.37 (dd, J =9.2, 0.8 Hz, 1H), 7.25- 7.18 (m, 3H), 6.49 (d, J = 6.4 Hz, 1H), 5.01(dd, J = 8.4, 3.2 Hz, 1H), 4.85 (dd, J = 3.6, 3.2 Hz, 1H), 3.90 (s, 3H),3.44-3.12 (m, 6H), 2.91-2.84 (m, 2H), 2.62 (s, 3H), 2.97-2.26 (m, 1H),2.16-2.00 (m, 4H), 1.93- 1.61 (m, 2H). I-175

453.3 2.20 7.80 (CDCl₃) δ 8.15 (t, J = 3.2 Hz, 1H), 7.31 (d, J = 8.8 Hz,1H), 7.10-7.01 (m, 3H), 6.31-6.27 (m, 1H), 4.45-4.42 (m, 1H), 4.26- 4.23(m, 1H), 3.85 (m, 3H), 3.44-3.20 (m, 3H), 3.02-2.91 (m, 1H), 2.58- 2.39(m, 2H), 2.17-2.10 (m, 1H), 1.92-1.83 (m, 5H), 1.65-1.59 (m, 2H),0.76-0.63 (m, 3H), 0.57- 0.47 (m, 1H). I-187

391.4 1.51 6.95 (CDCl₃) δ 8.43 (dd, J = 4.8, 1.2 Hz, 1H), 7.42 (dd, J =7.2, 1.6 Hz, 1H), 7.31 (d, J = 9.2 Hz, 1H), 7.13 (dd, J = 9.2, 7.2 Hz,1H), 7.05 (dd, J = 7.6, 4.8 Hz, 1H), 6.27 (dd, J = 7.2, 1.2 Hz, 1H),4.24-4.21 (m, 1H), 4.18-4.15 (m, 1H), 3.14 (s, 4H), 2.65 (s, 4H), 2.40(s, 6H), 2.25-2.22 (m, 1H), 2.15-2.12 (m, 1H), 1.85-1.76 (m, 3H), 1.72-1.65 (m, 2H). I-191

407.1 1.08 6.85 (CDCl₃) δ 8.18 (t, J = 6.0 Hz, 1H). 7.53 (s, 1H), 7.31(d, J = 9.2 Hz, 1H), 7.15- 7.11 (m, 3H), 6.26 (dd, J = 7.2, 0.8 Hz, 1H),4.46 (dd, J = 1.6, 1.2 Hz, 1H), 4.19 (d, J = 10.0 Hz, 1H), 3.85 (s, 3H),3.13 (s, 4H), 2.65 (s, 4H), 2.39 (s, 3H), 2.24- 2.20 (m, 1H), 2.11-2.07(m, 1H), 1.95-1.92 (m, 1H), 1.86-1.76 (m, 3H), 1.70-1.63 (m, 1H). I-152

405.3 1.20 7.20 (CDCl₃) 8.46 (s, 1H), 8.05 (d, J = 6.4 Hz, 1H), 7.51 (d,J = 9.2 Hz, 1H), 7.40 (d, J = 6.8 Hz, 1H), 7.11- 6.99 (m, 2H), 6.75-6.72(m, 1H), 3.79-3.55 (m, 2H), 3.54-3.35 (m, 2H), 3.12-2.91 (m, 2H), 2.87-2.76 (m, 2H), 2.59-2.49 (m, 2H), 2.42 (s, 3H), 2.33-2.20 (m, 2H), 2.16(s, 6H), 2.10-2.00 (m, 1H), 1.94-1.87 (m, 1H), 1.75- 1.55 (m, 2H). I-156

417.4 2.14 7.59 (CDCl₃) δ 8.49 (d, J = 3.6 Hz, 1H), 7.64 (s, 1H), 7.43(d, J = 7.6 Hz, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.13 (dd, J = 8.8, 7.2Hz, 1H), 7.06 (dd, J = 7.6, 4.8 Hz, 1H), 6.24 (d, J = 7.6 Hz, 1H), 3.58(dd, J = 11.2, 2.8 Hz, 1H), 3.47- 3.43 (m, 1H), 3.23-3.18 (m, 2H), 3.08(brs, 2H), 2.94-2.91 (m, 2H), 2.51- 2.46 (m, 3H), 2.05-1.96 (m, 3H),1.91 (s, 3H), 1.72-1.58 (m, 3H), 0.79- 0.65 (m, 4H). I-159

437.3 2.30 7.20 (CDCl₃) δ 8.61 (dd, J = 4.4, 1.6 Hz, 1H), 7.67 (s, 1H),7.65 (d, J = 1.6 Hz, 1H), 7.30 (d, J = 8.8 Hz, 1H), 7.15-7.11 (m, 2H),6.23 (dd, J = 7.2, 1.2 Hz, 1H), 3.96 (dd, J = 11.2, 2.8 Hz, 1H), 3.51(dd, J = 11.2, 3.2 Hz, 1H), 3.23- 3.18 (m, 2H), 3.07 (brs, 2H),3.00-2.95 (m, 2H), 2.10-1.96 (m, 4H), 1.93 (s, 3H), 1.86-1.61 (m, 3H),0.78-0.64 (m, 4H). I-160

437.3 2.25 8.00 (CDCl₃) δ 8.49-8.48 (d, J = 3.6 Hz, 1H), 7.64 (dd, J =8, 7.2 Hz, 1H), 7.55 (s, 1H), 7.32 (d, J = 8.8 Hz, 1H), 7.17-7.10 (m,2H), 6.27 (d, J = 7.2 Hz, 1H), 4.53 (dd, J = 11.2, 8.8 Hz, 1H),4.20-4.17 (m, 1H), 3.13 (s, 4H), 2.92 (brs, 2H), 2.44 (s, 3H), 2.22-2.19 (m, 1H), 2.03-2.00 (m, 1H), 1.89-1.76 (m, 2H), 1.59-1.52 (m, 1H),1.26 (brs, 2H), 0.88-0.84 (m, 2H), 0.60-0.57 (m, 2H). I-167

457.3 1.77 8.67 (CDCl₃) δ 8.48 (d, J = 4.4 Hz, 1H), 7.63 (dd, J = 8.4,1.2 Hz, 1H), 7.34 (d, J = 8.8 Hz, 1H), 7.11-7.07 (m, 2H), 6.33 (t, J =9.6 Hz, 1H), 4.52-4.50 (m, 1H), 4.28-4.26 (m, 1H), 3.44- 3.36 (m, 2H),3.34 (dd, J = 39.2, 11.2 Hz, 1H), 3.01- 2.92 (m, 1H), 2.58-2.41 (m, 2H),2.13-1.88 (m, 3H), 1.54-1.26 (m, 3H), 0.74-0.49 (m, 4H). I-172

419.3 1.91 6.72 (CDCl₃) δ 8.18 (t, J = 2.8 Hz, 1H), 7.52 (s, 1H), 7.31(d, J = 9.2 Hz, 1H), 7.15- 7.11 (m, 3H), 6.21 (dd, J = 7.2, 0.8 Hz, 1H),4.46 (dd, J = 11.2, 2.4 Hz, 1H), 4.17 (dd, J = 11.2, 2.8 Hz, 1H), 3.85(s, 3H), 3.17-3.15 (m, 2H), 3.08 (brs, 2H), 2.92 (s, 2H), 2.24-2.21 (m,1H), 2.12-2.08 (m, 1H), 1.96- 1.92 (m, 6H), 0.73-0.72 (m, 2H), 0.65-0.63(m, 2H). I-186

417.3 2.15 7.74 (CDCl₃) δ 8.35 (d, J = 4.0 Hz, 1H), 7.73 (s, 1H), 7.55(d, J = 7.6 Hz, 1H), 7.22- 7.10 (m, 3H), 6.33 (d, J = 7.2 Hz, 1H),4.46-4.43 (m, 1H), 4.34-4.31 (m, 1H), 3.96 (s, 2H), 2.80-2.77 (m, 2H),2.50 (d, J = 10 Hz, 2H), 2.34 (s, 3H), 2.26 (s, 3H), 2.21-2.18 (m, 1H),2.11-2.03 (m, 1H), 1.94 (s, 6H), 1.87-1.80 (m, 1H), 1.63-1.48 (m, 2H).I-190

403.4 1.53 6.82 (CDCl₃) δ 8.42 (d, J = 4.4 Hz, 1H), 7.58 (s, 1H), 7.42(d, J = 7.2 Hz, 1 H), 7.32- 7.29 (m, 1H), 7.16-7.11 (m, 1H), 7.06 (dd, J= 7.6, 4.8 Hz, 1H), 6.23 (d, J = 7.2 Hz, 1H), 4.26 (d, J = 11.2 Hz, 1H),4.19 (d, J = 8.8 Hz, 1H) 3.18-3.15 (m, 2H), 3.16 (s, 2H), 2.93 (s, 2H),2.41-2.22 (m, 1H), 2.16-2.12 (m, 1H), 1.88- 1.82 (m, 4H), 1.55-1.52 (m,2H), 0.75-0.70 (m, 2H), 0.67-0.65 (m, 2 H). I-192

405.2 2.19 7.50 (CDCl₃) δ 8.40 (d, J = 3.6 Hz, 1H), 7.39-7.33 (m, 2H),7.03-6.99 (m, 2H), 6.36 (d, J = 6.8 Hz, 1H), 4.21-4.18 (m, 1H), 4.13-4.10 (m, 1H), 3.19-3.13 (m, 2H), 2.97-2.91 (m, 1H), 2.89-2.84 (m, 3H),2.79 (s, 3H), 2.43-2.38 (m, 8H), 2.15-2.13 (m, 1H), 1.98-1.94 (m, 2H),1.82- 1.81 (m, 2H), 1.68-1.62 (m, 1H). I-193

425.2 2.05 8.29 (DMSO-d₆) δ 8.54 (dd, J = 4.8, 1.2 Hz, 1H), 7.91 (dd, J= 8.0, 1.2 Hz, 1H), 7.35- 7.32 (m, , 1H), 7.26 (d, J = 8.4 Hz, 1H), 7.09(dd, J = 8.8 Hz, 7.6 Hz, 1H), 6.51 (d, J = 6.8 Hz, 1H), 4.39- 4.32 (m,1H), 4.07-4.00 (m, 1H), 3.13-3.08 (m, 2H), 2.84-2.77 (m, 4H), 2.73 (s,3H), 2.45-2.39 (m, 1H), 2.30-2.24 (m, 5H), 2.03-1.98 (m, 1H), 1.85- 1.81(m, 2H), 1.70-1.59 (m, 2H), 1.30-1.26 (m, 1H). I-208

370.4 1.54 7.21 (CDCl₃) δ 8.74 (dd, J = 2.2, 0.6 Hz, 1H), 8.63 (dd, J =5.2, 1.6 Hz, 1H), 8.33 (dd, J = 4.8, 1.2 Hz, 1H), 7.98-7.95 (m, 1H),7.85- 7.81 (m, 1H), 7.60 (d, J = 9.2 Hz, 1H), 7.44-7.40 (m, 1H), 7.30(dd, J = 7.8, 1.0 Hz, 1H), 7.15-7.11 (m, 1H), 6.96-6.92 (m, 1H),6.71-6.66 (m, 1H), 4.04- 3.94 (m, 2H), 2.70-2.59 (m, 1H), 2.26 (s, 3H),2.08-1.94 (m, 2H), 1.83- 1.66 (m, 2H), 1.61-1.52 (m, 2H). I-209

370.4 1.65 7.87 (CDCl₃) δ 9.11 (d, J = 6.8 Hz, 1H), 8.75 (d, J = 4.4 Hz,1H), 8.41 (d, J = 4.0 Hz, 1H), 7.84-7.82 (m, 2H), 7.66 (d, J = 8.8 Hz,1H), 7.40 (d, J = 7.6 Hz, 1H), 7.25-7.21 (m, 2H), 7.04-7.01 (m, 1H),6.80 (t, J = 6.8 Hz, 1H), 4.28-4.20 (m, 2H), 2.40 (s, 3 H), 2.21-2.15(m, 3H), 1.95- 1.68 (m, 4H). I-210

405.4 1.40 6.39 (CDCl₃) δ 8.43 (d, J = 3.6 Hz, 1H), 7.54 (s, 1H),7.43-7.37 (m, 2H), 7.16 (dd, J = 9.2, 7.2 Hz, 1H), 7.06 (dd, J = 7.6,4.8 Hz, 1H), 6.27-6.26 (m, 1H), 4.24-4.15 (m, 2H), 3.83 (s, 2H),3.51-3.49 (m, 2H), 3.43-3.40 (m, 2H), 3.07 (s, 3H), 2.40 (s, 3H), 2.27-2.24 (m, 1H), 2.15-2.11 (m, 1H), 1.86-1.80 (m, 2H), 1.67-1.61 (m, 2H).I-211

405.4 1.35 6.08 (CDCl₃) δ 8.43 (d, J = 4.0 Hz, 1H), 7.56 (t, J = 9.2 Hz,1H), 7.42 (d, J = 6.8 Hz, 2H), 7.21-7.17 (m, 1H), 7.07-7.04 (m, 1H),6.71 (d, J = 6.8 Hz, 1H), 4.22-4.14 (m, 2H), 3.73- 3.70 (m, 2H),3.48-3.38 (m, 1H), 3.33-3.23 (m, 1H), 2.93-2.80 (m, 2H), 2.45-2.39 (m,6H), 2.30- 2.10 (m, 3H), 1.73-1.58 (m, 3H). I-212

373.4 1.52 7.14 (CDCl₃) δ 8.40 (d, J = 4.4 Hz, 1H), 7.66-7.64 (m, 2H),7.40 (d, J = 7.6 Hz, 1H), 7.35 (s, 1H), 7.25- 7.21 (m, 1H), 7.06-7.03(m, 1H), 6.77 (d, J = 6.8 Hz, 1H), 6.56 (d, J = 1.6 Hz, 1H), 4.20-4.12(m, 2H), 3.77 (s, 3H), 2.37 (s, 3H), 2.23-2.21 (m, 1H), 2.13-2.00 (m,1H), 1.84- 1.80 (m, 2H), 1.69-1.57 (m, 2H). I-213

373.4 1.40 6.35 (CDCl₃) δ 8.40 (d, J = 4.4 Hz, 1H), 7.66-7.62 (m, 2H),7.41 (d, J = 7.2 Hz, 1H), 7.36-7.33 (m, 2H), 7.22 (dd, J = 8.8, 6.8 Hz,1H), 7.06-7.03 (m, 1H), 6.75 (d, J = 6.8 Hz, 1H), 4.21-4.10 (m, 2H),3.52 (s, 3H), 2.39 (s, 3H), 2.24- 2.21 (m, 1H), 2.13-2.09 (m, 1H),1.81-1.77 (m, 2H), 1.71-1.57 (m, 2H). I-214

370.2 1.48 6.94 (CDCl₃) δ 8.81 (dd, J = 4.4, 1.6 Hz, 2H), 8.40 (d, J =4.0 Hz, 1H), 7.62 (d, J = 7.6 Hz, 2H), 7.56 (dd, J = 4.4, 1.6 Hz, 2H),7.41 (d, J = 7.2 Hz, 1H), 7.25-7.23 (m, 1H), 7.06-7.03 (m, 1H),6.77-6.75 (m, 1H), 4.21-4.13 (m, 2H), 2.37 (s, 3H), 2.26-2.21 (m, 1H),2.13-2.09 (m, 1H), 1.84- 1.81 (m, 2H), 1.69-1.57 (m, 2H). I-215

370.2 1.50 7.00 (CDCl₃) δ 8.84 (d, J = 0.8 Hz, 1H), 8.75 (d, J = 4.8 Hz,1H), 8.39 (d, J = 4.4 Hz, 1H), 7.97 (d, J = 7.6 Hz, 1H), 7.61 (d, J =9.2 Hz, 1H), 7.55 (s, 1H), 7.48 (dd, J = 8.0, 4.8 Hz, 1H), 7.40 (d, J =7.6 Hz, 1H), 7.26-7.22 (m, 1H), 7.05- 7.02 (m, 1H), 6.72 (d, J = 7.2 Hz,1H), 4.20-4.13 (m, 2H), 2.46 (s, 3H), 2.26- 2.22 (m, 1H), 2.13-2.09 (m,1H), 1.84-1.77 (m, 2H), 1.71-1.57 (m, 2H). I-216

370.3 1.32 5.41 (CDCl₃) δ 8.80-8.78 (m, 1H), 8.40-8.39 (m, 1H), 8.32 (s,1H), 7.88-7.84 (m, 1H), 7.71 (d, J = 7.6 Hz, 1H), 7.63 (d, J = 8.8 Hz,1H), 7.40-7.37 (m, 2H), 7.26-7.22 (m, 1H), 7.04- 7.00 (m, 2H), 4.22-4.16(m, 2H), 2.38 (s, 3H), 2.28-2.25 (m, 1H), 2.13- 2.11 (m, 1H), 1.84-1.66(m, 3H), 1.26-1.24 (m, 1H). I-217

388.3 1.63 7.90 ¹H NMR (400 MHz, CD₃OD): δ 8.42-8.39 (m, 2H), 7.96 (s,1H), 7.63 (d, J = 6.80 Hz, 1H), 7.53- 7.51 (m, 2H), 7.32-7.27 (m, 1H),7.25-7.22 (m, 1H), 6.93-6.89 (m, 1H), 6.78 (s, 1H), 4.13 (dd, J = 11.2,2.80 Hz, 1H), 3.95- 3.91 (m, 1H), 2.87-2.67 (m, 3H), 2.65-2.56 (m, 4H),2.03-1.97 (m, 4H), 1.81-1.66 (m, 2H). I-218

437.3 1.65 8.17 ¹H NMR (400 MHz, CD₃OD) δ: 8.38 (d, J = 4.0 Hz, 1H),8.05 (d, J = 6.4 Hz, 1H), 7.63 (s, 1H), 7.54 (d, J = 7.2 Hz, 1H), 7.45(d, J = 8.0 Hz, 1H), 7.37- 7.34 (m, 2H), 7.26-7.22 (m, 1H), 7.19-7.13(m, 3H), 6.85-6.80 (m, 1H), 4.17-4.12 (m, 1H), 3.95- 3.91 (m, 1H),2.95-2.86 (m, 2H), 2.68-2.62 (m, 1H), 2.58 (s, 3H), 2.45- 2.41 (m, 1H),2.06-1.97 (m, 4H), 1.81-1.68 (m, 2H).

Example 27: Synthesis of I-207

The Synthesis of X4-618-9

To a solution of X4-618-7 (2.0 g, 7.4 mmol), X4-618-8 (1.4 g, 11.3mmol), Cs2CO3 (4.8 g, 14.7 mmol) in 1,2-dimethoxyethane/H₂O (40 mL/8mL), were added (Cy)₃P (416.1 mg, 1.5 mmol) and Pd(dppf)Cl₂.CH₂C12(606.2 mg, 0.7 mmol) under N2 atmosphere. Then the solution was stirredat 95° C. overnight and the mixture was cooled to room temperature,filtered and extracted with DCM (40 mL×3). The combined organic layerswere washed with brine (20 mL×2), dried over anhydrous Na₂SO₄, filteredand concentrated in vacuum. The residue was purified by columnchromatography to give X4-618-9 (1.0 g, yield: 50%) as an off-whitesolid. LCMS (Agilent LCMS 1200-6120, Column: Waters X-Bridge C18 (30mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 2.0 mL/min;Mobile Phase: from 90% [water+10 mM NH₄HCO₃] and 10% [CH₃CN] to 5%[water+10 mM NH₄HCO₃] and 95% [CH₃CN] in 0.5 min, then under thiscondition for 1.5 min, finally changed to 90% [water+10 mM NH₄HCO₃] and10% [CH₃CN] in 0.1 min and under this condition for 0.5 min). Purity:88.73%, Rt=0.78 min; MS Calcd.: 267.3; MS Found: 268.2 [M+H]⁺.

The Synthesis of X4-618-10

To a solution of X4-618-9 (300 mg, 1.1 mmol), ethyl acetate (593.3 mg,6.7 mmol) in THF (5 mL) was added LiHMDS (1 M in THF, 3.4 mL, 3.4 mmol)and stirred at room temperature overnight. The reaction was quenchedwith 2 M HCl (50 mL) solution and washed with MTBE (3×10 mL). Then pHwas adjusted to 9 with 40% NaOH (aq.) and extracted with DCM (3×10 mL).The combined organic layers were washed with brine (10 mL×2), dried overanhydrous Na₂SO₄, filtered and concentrated in vacuum to give the crudeof X4-619-10 (300 mg, yield: 86%) as brown oil, which was used to thenext step without further purification. LCMS (Agilent LCMS 1200-6120,Column: Waters X-Bridge C18 (30 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 90% [water+10 mMNH₄HCO₃] and 10% [CH₃CN] to 5% [water+10 mM NH₄HCO₃] and 95% [CH₃CN] in0.5 min, then under this condition for 1.5 min, finally changed to 90%[water+10 mM NH₄HCO₃] and 10% [CH₃CN] in 0.1 min and under thiscondition for 0.5 min.). Purity: 90.84%, Rt=0.87 min; MS Calcd.: 309.3;MS Found: 310.2 [M+H]⁺.

The Synthesis of X4-618-11

To a solution of X4-618-10 (200 mg, 0.6 mmol), K₂CO₃ (82.8 mg, 0.6 mmol)in CH₃CN (10 ml) under Ar protection was added X4-618-3 (147 mg, 1.0mmol). The mixture was stirred at room temperature overnight. Then itwas poured into water (30 mL) and extracted with DCM (3×100 mL). Thecombined organic layers were washed with brine (10 mL×2), dried overanhydrous Na₂SO₄, filtered and concentrated in vacuum to give the crudeof X4-619-11 (90 mg, yield: 30%) as brown oil, which was used to thenext step without further purification. LCMS (Agilent LCMS 1200-6120,Column: Waters X-Bridge C18 (30 mm*4.6 mm*3.5 μm); Column Temperature:40° C.; Flow Rate: 2.0 mL/min; Mobile Phase: from 90% [water+10 mMNH₄HCO₃] and 10% [CH₃CN] to 5% [water+10 mM NH₄HCO₃] and 95% [CH₃CN] in0.5 min, then under this condition for 1.5 min, finally changed to 90%[water+10 mM NH₄HCO₃] and 10% [CH₃CN] in 0.1 min and under thiscondition for 0.5 min). Purity: 75.81%, Rt=1.04 min; MS Calcd.: 456.3;MS Found: 457.2 [M+H]⁺.

The Synthesis of X4-618-12

A solution of X4-618-11 (90 mg, 0.2 mmol) in conc. HCl solution (2 mL)was stirred at 100° C. for 2 h. Then it was concentrated in vacuo. Theresidue was dissolved in H₂O (10 mL), neutralized with 20% NaOH aq. toadjust to pH >9, and extracted with DCM (3×10 mL). The combined organiclayers were washed with brine (10 mL×2), dried over anhydrous Na₂SO₄,filtered and concentrated in vacuum to give crude X4-619-12 (70 mg,yield: 92%) as brown semi-solid, which was used to the next step withoutfurther purification. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (30 mm*4.6 mm*3.5 μm); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 90% [water+10 mM NH₄HCO₃] and 10%[CH₃CN] to 5% [water+10 mM NH₄HCO₃] and 95% [CH₃CN] in 0.5 min, thenunder this condition for 1.5 min, finally changed to 90% [water+10 mMNH₄HCO₃] and 10% [CH₃CN] in 0.1 min and under this condition for 0.5min.). Purity: 78.25%, Rt=0.96 min; MS Calcd.: 384.3; MS Found: 385.2[M+H]⁺.

The Synthesis of I-207

A solution of X4-618-12 (70 mg, 0.2 mmol), NH₄Br (53.5 mg, 0.55 mmol),AcOH (12.0 mg, 0.2 mmol), KOH (2.5 mg, 0.05 mmol), and NaBH₃CN (17.2 mg,0.3 mmol) in dry CH₃OH (5 ml) under Ar protection was stirred at roomtemperature overnight, then heated at reflux for 8 h. Then it was cooledto room temperature and quenched with 10 mL of H₂O, extracted with DCM(3×20 mL). The combined organic layers were washed with brine (10 mL×2),dried over anhydrous Na₂SO₄, filtered and concentrated in vacuum. Theresidue was purified by prep-HPLC to give I-207 (20 mg, yield: 30%) asan off-white solid. LCMS (Agilent LCMS 1200-6120, Column: WatersX-Bridge C18 (50 mm*4.6 mm*3.5 μm)); Column Temperature: 40° C.; FlowRate: 2.0 mL/min; Mobile Phase: from 95% [water+10 mM NH₄HCO₃] and 5%[CH₃CN] to 0% [water+10 mM NH₄HCO₃] and 100% [CH₃CN] in 1.6 min, thenunder this condition for 1.4 min, finally changed to 95% [water+10 mMNH₄HCO₃] and 5% [CH₃CN] in 0.1 min and under this condition for 0.7min.). Purity: 100.00%. Rt=1.53 min; MS Calcd.: 369.3; MS Found: 370.2[M+H]⁺. HPLC (Agilent HPLC 1200, Column: Waters X-Bridge C18 (150 mm*4.6mm*3.5 μm); Column Temperature: 40° C.; Flow Rate: 1.0 mL/min; MobilePhase: from 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] to 0% [water+10 mMNH₄HCO₃] and 100% [CH₃CN] in 10 min, then under this condition for 5min, finally changed to 95% [water+10 mM NH₄HCO₃] and 5% [CH₃CN] in 0.1min and under this condition for 5 min). Purity: 95.73%. Rt=7.12 min. ¹HNMR (400 MHz, CDCl₃) δ: 8.78 (d, J=5.6 Hz, 2H), 8.41 (d, J=4.0 Hz, 1H),8.18 (d, J=7.2 Hz, 1H), 7.68 (d, J=8.8 Hz, 1H), 7.52 (d, J=3.6 Hz, 2H),7.39 (d, J=7.2 Hz, 1H), 7.24 (d, J=8.0 Hz, 1H), 7.03 (dd, J=7.6, 5.2 Hz,1H), 6.80 (t, J=6.8 Hz, 1H), 4.12 (s, 2H), 2.36 (s, 3H), 2.16-2.06 (m,2H), 1.87-177 (m, 5H).

Example 28: REGA Screening Assay Intracellular CXCL-12-Induced CalciumMobilization Assay

Intracellular calcium mobilization induced by chemokines orchemokine-derived peptides were evaluated using a calcium responsivefluorescent probe and a FLIPR system. The CXCR-4 transfected U87 cellline (U87.CXCR4) cells were seeded in gelatine-coated black-wall 96-wellplates at 20,000 cells per well and incubated for 12 hours. Cells werethen loaded with the fluorescent calcium probe Fluo-2 acetoxymethyl at 4μM final concentration in assay buffer (Hanks' balanced salt solutionwith 20 mM HEPES buffer and 0.2% bovine serum albumin, pH 7.4) for 45min at 37° C. The intracellular calcium mobilization induced by theCXCL-12 (25-50 ng/mL) was then measured at 37° C. by monitoring thefluorescence as a function of time in all the wells simultaneously usinga fluorometric imaging plate reader (FLIPR Tetra, Molecular Devices).The test compounds were added 15 minutes before the addition of CXCL-12and monitored to see if compounds induced signals by themselves(agonistic properties).

Chemokine (CXCL12-AF647) Binding Inhibition Assay

Jurkat cells expressing CXCR4 were washed once with assay buffer (Hanks'balanced salt solution with 20 mM HEPES buffer and 0.2% bovine serumalbumin, pH 7.4) and then incubated for 15 min at room temperature withthe test compounds diluted in assay buffer at dose-dependentconcentrations. Subsequently, CXCL12-AF647 (25 ng/mL) was added to thecompound-incubated cells. The cells were incubated for 30 min at roomtemperature. Thereafter, the cells were washed twice in assay buffer,fixed in 1% paraformaldehyde in PBS, and analyzed on the FL4 channel ofa FACSCalibur flow cytometer equipped with a 635-nm red diode laser(Becton Dickinson, San Jose, Calif., USA).

The percentages of inhibition of CXCL12-AF647 binding were calculatedaccording to the formula: [1−((MFI−MFI_(NC))/(MFI_(PC)−MFI_(NC)))]×100where MFI is the mean fluorescence intensity of the cells incubated withCXCL12-AF647 in the presence of the inhibitor, MFI_(NC) is the meanfluorescence intensity measured in the negative control (i.e.,autofluorescence of unlabeled cells), and MFI_(PC) is the meanfluorescence intensity of the positive control (i.e., cells exposed toCXCL12-AF647 alone).

Results of Assays

Table 3 shows the activity of selected compounds of this invention inthe assays described above. The compound numbers correspond to thecompound numbers in Table 1. Compounds having an activity designated as“A” provided an IC₅₀ of 0.01 to 100 nM; compounds having an activitydesignated as “B” provided an IC₅₀ of >100 nm to <1 μM; and compoundshaving an activity designated as “C” provided an IC₅₀ of 1 μM orgreater.

TABLE 3 Inhibition of Ca²⁺ Signalling and Inhibition of CXCL12 BindingIC₅₀ CXCL-12 Ca2+ flux IC₅₀ CXCL-12 Compound U87.CXCR4+ binding Jurkat #(nM) (nM)  I-1 A A  I-2 B B  I-3 A A  I-4 A A  I-5 B A  I-6 A B  I-7 C B I-8 A A  I-9 A A  I-10 A A  I-11 A A  I-12 A A  I-13 A A  I-14 B B I-15 A A  I-16 A A  I-17 A A  I-18 A A  I-19 A A  I-20 B A  I-21 A A I-25 A A  I-26 B A  I-27 A A  I-28 B A  I-29 B A  I-34 A A  I-37 A A I-38 A A  I-39 B A  I-40 A A  I-45 C B  I-46 A A  I-47 A A  I-48 B A I-49 A A  I-50 A A  I-51 B A  I-52 C B  I-53 B A  I-54 A A  I-55 C A I-56 A A  I-57 A A  I-58 C B  I-59 C B  I-60 A A  I-61 A A  I-62 C B I-63 B A  I-66 B A  I-67 B A  I-68 A A  I-69 A A  I-73 C B  I-74 C B I-75 C B  I-76 A A  I-77 A A  I-78 A A  I-79 A A  I-80 B A  I-81 B A I-82 A A  I-83 B A  I-85 A A  I-86 A A  I-87 A A  I-88 A A  I-89 A A I-91 C B  I-92 A A  I-93 B A I-104 B A I-105 C B I-106 A A I-109 B AI-110 A A I-111 C B I-112 B A I-113 A A I-114 A A I-115 A A I-116 A AI-117 A A I-119 C A I-123 C A I-125 B A I-126 A A I-127 A A I-132 B AI-134 A A I-135 A A I-136 A A I-137 B A I-138 A A I-139 B A I-140 A AI-141 B A I-142 A A I-146 B A I-147 B A I-149 A A I-150 C A I-151 B AI-152 A A I-154 C B I-156 A A I-159 A A I-160 C B I-164 A A I-166 C BI-167 C B I-172 C C I-173 C B I-175 C C I-179 A A I-180 A A I-181 A AI-182 C A I-186 A A I-187 A A I-188 A A I-189 B A I-190 C B I-191 B AI-192 A A I-193 A A I-207 C C I-208 C C I-209 — C I-210 C C I-211 C BI-212 C C I-213 C C I-214 C C I-215 C C I-216 C C I-219 C C

Example 28: Caco-2 Permeability Assay Assay Procedure

The goal of this assay was to evaluate the intestinal absorptionpotential of drug candidates using Caco-2 cell lines.

Experimental Procedure

1. Prewarm Prewarm HBSS Buffer in 37° C. water bath2. Sonicate Take compounds from −20° C., sonicate for a few minutes (noless than 1 minute)3. Solution preparation

Donor Solution Buffer For A-to-B Direction:

HBSS buffer with 0.3% DMSO and 5 μM LY: add 150 μL DMSO and 50 μL LY (5mM) into 50 ml HBSS buffer (pH 7.4).HBSS buffer with 0.1% DMSO and 5 μM LY: add 50 μL DMSO and 50 μL LY (5mM) into 50 mL HBSS buffer (pH 7.4).

For B-to-A Direction:

HBSS buffer with 0.3% DMSO: add 150 μL DMSO into 50 ml HBSS buffer (pH7.4).HBSS buffer with 0.1% DMSO: add 50 μL DMSO into 50 ml HBSS buffer (pH7.4).

Receiver Solution Buffer: For A-to-B Direction:

Prepare HBSS buffer with 0.4% DMSO: add 200 μL DMSO into 50 ml HBSSbuffer (pH 7.4).

For B-to-A Direction:

Prepare HBSS buffer with 0.4% DMSO and 5 μM LY: add 200 μL DMSO and 50μL LY (5 mM) into 50 ml HBSS buffer (pH 7.4).

TABLE 4 Preparation of Test Solutions Stock Solution (in DMSO) FinalDMSO Compound Test cpd Verapamil apical Buffer basolateral bufferconcentration Erythromycin + A-to-B dosing solution 10 mM 3 μL — 0.1%DMSO — 0.4% Metroprolol + HBSS + LY 3 mL Atenolol A-to-B Receiversolution — — — 0.4% DMSO HBSS 0.4% B-to-A dosing solution 10 mM 3 μL — —0.1% DMSO 0.4% HBSS 3 mL B-to-A Receiver solution — — 0.4% DMSO — 0.4%HBSS + LY cpds A-to-B dosing solution 10 mM 3 μL — 0.3% DMSO — 0.4%HBSS + LY 3 mL A-to-B Receiver solution — — — 0.4% DMSO HBSS 0.4% B-to-Adosing solution 10 mM 3 μL — — 0.3% DMSO 0.4% HBSS 3 mL B-to-A Receiversolution — — 0.4% DMSO — 0.4% HBSS + LY4. Measure TEER Take cell culture plate out of incubator, wash the cellmonolayers with HBSS buffer, and then measure TEER values at Rmtemperature.5. Centrifuge Centrifuge the compound solution (from step 3) at 4000 rpmfor 5 min before loading to donor chambers.6. Dosing Add solution based on the volumes listed in the followingtable (make sure to take extra 100 μL of donor sample for TO as Backup).

TABLE 5 Dosing Parameters Position Transport Direction Volume addedFinal volume Apical A-B (Donor chamber) 600 μL of A-to-B dosing solution(100 μL for LY 400 μL measurement and 100 μL for Backup) Basolateral A-B(Receiver chamber) 800 μL 0.4% DMSO HBSS 800 μL Basolateral B-A (Donorchamber) 900 μL B-to-A dosing solution (100 μL for 800 μL Backup) ApicalB-A (Receiver chamber) 500 μL 0.4% DMSO HBSS+ LY (100 μL for LY 400 μLmeasurement)7. Apical LYTO samples To determine LY concentration in the apicalchamber, take 100 μL sample from apical chambers into an opaque platefor LYTO.8. Prewarm Prewarm apical and basolateral plates at 37° C. for about 5min, then begin transport by placing the apical plate onto basolateralplate.9. Incubation Keep the plates in incubator at 37° C. for 90 min.10. Standard Curve preparation

Prepare 20× Solution

For 300 μM compound solution, add 6 μL of compound stock solution into192 μL of MeOH/H₂O (1:1).

Prepare Working Solution in MeOH/H₂O (1:1)

TABLE 6 Solutions for Standard Curve Preparation MeOH/H₂O Final Compoundsolution (μM) Solution (μL) (μL) solution (μM) 300 100 400 → 60 60 100200 → 20 20 100 400 → 4 4 100 400 → 0.8 0.8 100 300 → 0.2 0.2 100 100 →0.1

Prepare 1× Solution

3 μL (20×)+57 μL of 0.4% DMSO HBSS+60 μL ACN with IS (Osalmid orImipramine)—120 μL (1×)11. Transport termination Separate the apical plate from the basolateralplate after 90-min incubation.12. Measure LY Take 100 μL samples from basolateral plate to an opaqueplate as LYT90.13. Measure LY concentrations for LYTO and LYT90 by Fluorometer (atexcitation of 485 nm/emission of 535 nm).14. Sample preparation for LC-MS/NIS Donor samples (1:10 diluted): 6 μLof donor sample+54 μL 0.4% DMSO HBSS+60 μL ACN with IS (Osalmid orImipramine) Receiver sample: 60 μL of receiver sample+60 μL ACN with IS(Osalmid or Imipramine)

TABLE 7 Bioanalytical Conditions Detection LC-MS/MS-014(API4000) methodMatrix HBSS Internal Osalmid or Imipramine standard (s) MS conditionsPositive ion, ESI Mobile phase A: H₂O - 0.025% FA-1 mM NH₄OAC B: MeOH-0.025% FA-1 mM NH₄OAC Column Ultimate-XB-C18 (2.1 × 50 mm, 5 μm) LCconditions 0.60 mL/min Time (min) Pump B (%) 0.2  2 0.4 98 1.40 98 1.41 2 2.50 stop Detection & Analyte Retention time Analyte Mass RT IS MassRanges (RT) compound Ranges (Da) (min) (Da) IS RT (min) Erythromycin734.300/158.000 Da 0.90 281.100/193.100 Da 0.91 Metoprolol268.100/133.100 Da 0.85 281.100/193.100 Da 0.91 Atenolol 267.000/145.100Da 0.78 281.100/193.100 Da 0.91 I-11 435.299/417.186 Da 0.85281.100/193.100 Da 1.00 I-17 484.474/270.000 Da 0.87 281.100/193.100 Da1.00 I-13 419.231/175.097 Da 0.85 281.100/193.100 Da 1.00 I-14473.400/175.104 Da 1.01 281.100/193.100 Da 1.01 I-15 433.163/144.300 Da0.86 281.100/193.100 Da 1.02 I-16 449.400/157.028 Da 1.25230.100/121.200 Da 1.36

Results

Study details: Test concentration 10 μM

Reference compounds: Erythromycin, Metoprolol, Atenolol, Lucifer YellowTest systems: Caco-2/HBSS solutionIncubation conditions: 0, 90 min at 37° C.Sample size: Duplicates (n=2)Bioanalytical method: LC-MS/MS

Calculations

Transepithelial electrical resistance (TEER)=(Resistancesample−Resistance blank)×Effective Membrane Area

Lucifer Yellow permeability:Papp=(VA/(Area×time))×([RFU]accepter−[RFU]blank)/(([RFU]initial,donor−[RFU]blank)×DilutionFactor)×100

Drug permeability:Papp=(VA/(Area×time))×([drug]accepter/(([drug]initial,donor)×DilutionFactor)

Where VA is the volume in the acceptor well, area is the surface area ofthe membrane and time is the total transport time in seconds.

For Millicell-24 Cell Culture Plates: surface area of the membrane=0.7cm², VA=0.8 mL (A-to-B) or 0.4 mL (B-to-A)

Results

TEER value of Caco-2 monolayers from randomly selected wells was 357±29Ω·cm² (Mean±SD). Note: Cell monolayer is used if TEER value >100 Ω·cm².

Comments:

1. Papp values were calculated based on calculated concentrations.2. Most of the Caco-2 monolayers applied in this assay showed intacttight junctions as indicated by TEER values and low permeability forLucifer Yellow, a low permeability control (data not shown).3. Metoprolol, a high permeability control, showed both A-to-B andB-to-A permeability >10×10⁻⁶ cm/sec in Caco-2 cells. Atenolol, a lowpermeable control, showed both A-to-B and B-to-A permeability less than5×10⁻⁶ cm/sec in Caco-2 cells. Erythromycin, an efflux substrate, gavean efflux ratio higher than 116.11 in Caco-2 cells.4. As summarized in Table 8, compounds showing permeability <5×10⁻⁶cm/sec suggest low permeability; compounds showing permeability 5 to10×10⁻⁶ cm/sec suggest moderate permeability in A-to-B direction;compounds showing permeability >10×10⁻⁶ cm/sec suggest highpermeability.

Permeability results are shown in Table 8 for selected compounds of theinvention. The compound numbers correspond to the compound numbers inTable 1. Compounds having a ratio designated as “A” provided a ratio of0.1 to 10; compounds having a ratio designated as “B” provided a ratioof >10 to <30; and compounds having a ratio designated as “C” provided aratio of 30 or greater.

TABLE 8 Caco-Papp Permeability for Selected Compounds Compound Caco-Papp(A-B) Caco-Papp (B-A) Efflux ratio # (10⁻⁶ cm/sec) (10⁻⁶ cm/sec)(PB-A/PA-B)  I-1 A B A  I-3 A A A  I-4 A B C  I-6 B B A  I-9 B B A  I-11A B A  I-12 A B A  I-13 A B A  I-14 B B A  I-15 A B A  I-16 B B A  I-17A A A  I-18 A A A  I-19 A A A  I-20 A A A  I-21 A B A  I-25 A B C  I-27A A B  I-28 A B C  I-29 A B A  I-34 B A A  I-37 A B A  I-38 A C C  I-39A C B  I-40 A C C  I-45 A A A  I-46 A A A  I-47 A A A  I-48 A B A  I-49A B A  I-50 A B B  I-51 A B A  I-52 A C B  I-53 A B A  I-54 A A C  I-55A A A  I-56 A B A  I-57 A B A  I-58 B B A  I-60 A B A  I-61 A B A  I-62A B C  I-63 A B B  I-66 B B A  I-67 A A A  I-68 A B B  I-69 A B A  I-73B B A  I-75 A B A  I-76 A B B  I-77 B C A  I-78 A B C  I-79 A B A  I-80A A A  I-81 B B A  I-82 A B A  I-83 A B B  I-85 A B A  I-86 A B B  I-87A B B  I-88 A A A  I-89 A A A  I-91 A A A  I-92 A A A  I-93 B A A I-104A B C I-105 A B C I-106 A B A I-109 A B C I-110 A B C I-111 A A A I-112B B A I-113 A A A I-114 A A A I-115 A A A I-116 A B A I-117 B B A I-123A A A I-125 A A A I-126 A B B I-127 A A A I-132 A A A I-134 B B A I-135A B A I-136 A B B I-137 A A A I-138 B B A I-139 A C C I-140 A B A I-141A B A I-142 A B A I-146 B B A I-147 B B A I-149 B B A I-150 A B A I-151B B A I-152 A B A I-154 A B A I-156 A B A I-159 B B A I-160 B B A I-164B B A I-166 B B A I-167 A A A I-172 A C C I-173 A B A I-175 A B A I-179A B A I-180 B B A I-181 A B A I-182 B B A I-186 A B A I-187 A B A I-188B B A I-189 B B A I-190 A C B I-191 A C B I-192 A B A I-193 B B A I-207B C A I-208 B B A I-210 A B C

Example 29: Pharmacokinetics and Brain Penetration Experiment toDetermine Brain and Plasma Concentrations of Compounds after IVAdministration to Male CD1 Mice or Male SD Rat Mouse Study

In-Life Summary:

The study design (9 animals) consisted of administrating the drug (IV: 3mg/kg (5 mL/kg) via tail vein injection) and collecting samples atterminal bleeding for plasma and brain at 0.083, 0.5 and 1 h. The bloodcollection was performed as follows: the animal was restrained manuallyand approximately 150 μL blood/time point was collected into adipotassium EDTA tube via retro orbital puncture under anesthesia withisoflurane. The blood sample was put on ice and centrifuged to obtain aplasma sample (2000 g, 5 min under 4° C.) within 15 minutes. The braincollection was performed as follows: a mid-line incision was made in theanimal's scalp and the skin was retracted. Using small bone cutters androngeurs, the skull overlying the brain was removed. The brain wasremoved using a spatula and the brain rinsed with cold saline. The brainwas placed in screw-top tubes, and the tubes were stored at −70° C.until analysis. The IV dosing solution was prepared in 50 mM citratebuffer (pH 4.0) at 0.6 mg/mL.

Plasma Sample Preparation:

An aliquot of 30 μL sample was added to 150 μL MeCN containing 50 ng/mLIS (Dexamethasone). The mixture was vortexed for 5 min and centrifugedat 14,000 rpm for 5 min. An aliquot of 5 μL supernatant was injected forLC-MS/MS analysis.

Brain Sample Preparation:

An aliquot of 30 μL brain homogenate (brain:PBS=1:3, w/v) sample wasadded to 150 μL IS in ACN (Dexamethasone, 50 ng/mL). The mixture wasvortexed for 5 min and centrifuged at 14,000 rpm for 5 min. An aliquotof 5 μL supernatant was injected for LC-MS/MS analysis.

Analytical Method:

The sample analysis was performed on LCMS/MS-003 (API4000, triplequadruple) under the following conditions: positive ion, ESI, MRMdetection using dexamethasone as internal standard. HPLC conditions:mobile phase A: H₂O (0.025% formic acid (FA) with 1 mM NH₄OAc); mobilephase B: MeOH (0.025% FA with 1 mM NH₄₀Ac) on Waters X-Bridge C18(2.1×50 mm, 2.5 μm) column at 60° C.

Rat Study

In-Life Summary:

The study design consisted of 2 groups (24 animals and 18 animals) andadministrating the drug [IV: 3 mg/kg (1.5 mL/kg) via foot dorsal vein],[PO: 10 mg/kg (5 mL/kg) via oral gavage] and collecting samples atterminal bleeding for plasma, brain and CSF at 0.25, 0.5, 1, 4, 8 and 24hr. The IV and PO dosing solutions were prepared in 50 mM citrate buffer(pH 4.0) at 2 mg/mL. The blood collection was performed as follows: theanimal was restrained manually at the designated time points,approximately 150 μL of blood sample was collected via cardiac puncturevein into EDTA-2K tubes. The blood samples were maintained in wet icefirst and centrifuged to obtain plasma (2000 g, 4° C., 5 min) within 15minutes post sampling. The brain collection was performed as follows: amid-line incision was made in the animal's scalp and the skin wasretracted. Using small bone cutters and rongeurs, the skull overlyingthe brain was removed. The brain was removed using a spatula and rinsedwith cold saline. The brain was placed in screw-top tubes and thenstored under −70° C. until analysis. The CSF collection was performed asfollows: the animal was euthanized under deep anesthesia with air bubbletail vein injection. The CSF was collected by direct puncture ofbutterfly needle into the cisterna magna, using the occipital bone andthe wings of the atlas as landmarks. A piece of white paper was used asa background to monitor color change in the sample just above the needleduring collection. Upon observation of color change, the PE tubing wasquickly clamped off above the color change and cut just above theclamped site. The clear sample was drawn into the syringe.

Plasma Samples Preparation:

An aliquot of 30 μL sample was added with 100 MeCN containing 100 ng/mLIS (Dexamethasone). The mixture was vortexed for 10 min and centrifugedat 5800 rpm for 10 min. An aliquot of 40 μL supernatant was added with40 μl H₂O and the mixture was vortexed for 5 min. An aliquot of 2 μLsupernatant was injected for LC-MS/MS analysis.

Brain Samples Preparation:

The sample was homogenized with 3 volumes (v/w) of PBS. An aliquot of 30μL sample was added with 100 μL ACN containing 100 ng/mL IS(Dexamethasone). The mixture was vortexed for 10 min and centrifuged at5800 rpm for 10 min. An aliquot of 40 μL supernatant was added with 40μl H₂O and the mixture was vortexed for 5 min. An aliquot of 2 μLsupernatant was injected for LC-MS/MS analysis.

CSF Samples Preparation:

An aliquot of 10 μL sample was added with 10 μL MeOH/H₂O (1/1) and 404,ACN containing 200 ng/mL IS (Dexamethasone) 120 μL H₂O. The mixture wasvortexed for 5 min. An aliquot of 2 μL supernatant was injected forLC-MS/MS analysis.

Analytical Method:

The sample analysis was performed on UPLC-MS/MS-02 (Triple Quad™ 4000)under the following conditions: positive ion, ESI, MRM detection usingdexamethasone as internal standard. HPLC conditions: mobile phase A:H₂O-0.1% FA, mobile phase B: MeCN-0.1% FA on ACQUITY UPLC HSS T3 (2.1×50mm, 1.8 μm) column at 60° C.

TABLE 9 Mouse and Rat Brain Uptake Assay Results - 0-1 h, 3 Time PointsMouse (MBUA, 3 mg/kg, IV) Cl Fu (p) Fu (b) AUClast AUClast Kp, Compound# (L/hr/kg) t1/2 % % (p) (b) Kp uu I-6  2.9 3.6 531 326 0.61 I-18  14.70.344 5.4 0.2 175 2063 11.78 0.43 I-19  13.4 0.363 2.8 0.1 191 1690 8.840.32 I-20  10.4 0.374 2.1 0.1 241 1515 6.29 0.30 I-114 7.01 0.72 2.8 0.3267 205 0.76 0.08 I-146 0.433 6.02 5.3 0.6 884 1643 1.86 0.21 I-149 12.40.367 9.9 2.6 203 295 1.45 0.38 I-187 7.62 0.354 24.9 7.1 335 76.5 0.220.05 Rat (3 mg/kg IV; 10 mg/kg PO) Cl AUClast AUClast Kp, Compound #(L/hr/kg) t1/2 F % Fu (p) Fu (b) (p) (b) Kp uu I-146 1.46 7.89 70.1 70.7 4276 7800 1.82 0.18 I-149 7.63 6.7 37.9 20 3.8 457 3820 8.4 1.6I-188 4.46 6.87 36.9 30.8 5.7 791 4472 5.65 1.04 I-189 4.13 8.14 25.424.8 6.1 570 2296 4.03 0.99 The following abbreviations are used inTable 9: Cl: Clearance (L/hr/kg) t_(1/2): half life (in hours) Fu (p) %:Fraction of drug unbound to plasma proteins (%) Fu (b) %: Fraction ofdrug unbound to brain proteins (%) F %: oral bioavailability (the totalnumber for protein bound and free fraction) AUC last(p): Total areaunder the plasma drug concentration-time curve (time zero to 1 hr afterdrug administration) (hr*ng/mL) AUC last(b): Total area under the braindrug concentration-time curve (time zero to 1 hr after drugadministration) (hr*ng/mL) Kp: brain/plasma drug concentration ratio(AUC last(b)/AUC last(p)) Kp uu: unbound brain/unbound plasma drugconcentration ratio (calculated as follows: Fu (b)*AUC last (b)/Fu(p)*AUC last (p))

A similar experiment was performed with 4 time points taken from 0-24 h.The results are shown below in Table 10.

TABLE 10 Mouse Brain Uptake Assay Results - 0-24 h, 4 Time Points Mouse(MBUA, 3 mg/kg, IV) Cl Fu (p) Fu (b) AUClast AUClast Kp, Compound #(L/hr/kg) t1/2 % % (P) (b) Kp uu I-6  2.9 3.6 1037 3173 3.05 I-115 2.123.96 2.6 0.5 1402 26886 19.17 3.68 I-117 2.01 3.56 3.9 0.6 1480 3229721.82 3.35 I-146 0.433 6.02 5.3 0.6 6587 34840 5.29 0.6 I-187 1.72 4.7224.9 7.1 1715 1358 0.79 0.22 I-193 1.32 6.42 4.7 0.4 2131 13673 6.410.54

Example 30: MTD and Pharmacokinetics and Brain Penetration Experiment toDetermine Brain and Plasma Concentration of Compounds After POAdministration to Male C57BL/6 Mice

In-Life Summary:

The study was designed with 2 groups (18 animals and 24 animals)consists of administrating the drug [P0-50, 100, 150, 225, 300 mg/kg viaoral gavage] and collecting samples at terminal bleeding for plasma,brain and CSF at 0.25, 0.5, 1, 4, 8, and 24 hr. All PO dosing solutionswere prepared in 50 mM citrate buffer (pH 4.0).

TABLE 11 Administration of Compounds Schedule for Two Test Groups Group1: Single administration: PO: 50 mg/kg (10 mL/kg) via oral gavage (N =18) Group 2: multiple administrations: PO-day1: 50 mg/kg (10 mL/kg) viaoral gavage (N = 24) PO-day2: 100 mg/kg (10 mL/kg) via oral gavage (N =24) PO-day3: 150 mg/kg (10 mL/kg) via oral gavage (N = 24) PO-day4: 225mg/kg (10 mL/kg) via oral gavage (N = 24) PO-day5: 300 mg/kg (10 mL/kg)via oral gavage (N = 24)

The blood collection was performed as follows: the animal was restrainedmanually at the designated time points, approximately 500 μL of bloodsample was collected via cardiac puncture vein into EDTA-2K tubes. Thewhole blood needed to be divided into two parts; one part was placed inthe tube containing EDTA-2K for plasma generation and the other was usedfor the hematology assay, respectively. The blood samples for plasmageneration were maintained in wet ice first and centrifuged to obtainplasma (2000 g, 4° C., 5 min) within 15 minutes post sampling. The braincollection was performed as follows: a mid-line incision was made in theanimal's scalp and the skin was retracted. Using small bone cutters androngeurs, the skull overlying the brain was removed. The brain wasremoved using a spatula and rinsed with cold saline. The brain wasplaced in screw-top tubes, and then stored at −70° C. until analysis.The CSF collection was performed as follows: a mid line incision wasmade on the neck. The muscle under the skin was cut to expose thecisterna magna. The cisterna magna was penetrated with the sharp end ofone capillary (Burn one end of capillary to make it sharp). The CSF wassucked spontaneously into the capillary.

Plasma Sample Preparation:

An aliquot of 30 μL sample was added with 100 μL MeCN containing 100ng/mL IS (Dexamethasone). The mixture was vortexed for 10 min andcentrifuged at 5800 rpm for 10 min. An aliquot of 40 μL supernatant wasadded to 40 μL H₂O and the mixture was vortexed for 5 min. An aliquot of2 μL supernatant was injected for LC-MS/MS analysis.

Brain Sample Preparation:

An aliquot of 30 μL brain homogenate (brain:PBS=1:3, w/v) sample wasadded to 100 μL MeCN containing 100 ng/mL IS (Dexamethasone). Themixture was vortexed for 10 min and centrifuged at 5800 rpm for 10 min.An aliquot of 40 μL supernatant was added to 40 μL H₂O and the mixturewas vortexed for 5 min. An aliquot of 2 μL supernatant was injected forLC-MS/MS analysis.

CSF Samples Preparation:

An aliquot of 3 μL sample was added to a mixture of 6 μL CSF, 9 μLMeOH/H₂O (1/1), 40 μL MeCN containing 200 ng/mL IS (Dexamethasone), and116 μL H₂O. The mixture was vortexed for 5 min. An aliquot of 4 μL wasinjected for LC-MS/MS analysis.

Analytical Method:

The sample analysis was performed on UPLC-MS/MS-02 (Triple Quad™ 4000)under the following conditions: positive ion, ESI, MRM detection usingdexamethasone as internal standard. HPLC conditions: mobile phase A:H₂O-0.1% formic acid, mobile phase B: MeCN-0.1% formic acid on: ACQUITYUPLC HSS T3 (2.1×50 mm, 1.8 μm) at 60° C.

TABLE 12 Mouse MTD For Single Administration Group Mouse (MTD mouse,D1-50 mg/kg, PO) Cl Fu (p) Fu (b) AUClast AUClast Kp, (L/hr/kg) t1/2 % %(p) (b) Kp uu I-188 1-24 14.3 5.4 23496 58553 2.49 0.94

TABLE 13 Mouse MTD For Multiple Administration Group Mouse (MTD mouse,D5-300 mg/kg, PO) Cl Fu Fu AUClast AUClast Kp, (L/hr/kg) t1/2 (p) (b)(p) (b) Kp uu I-188 4-72 14.3 5.4 76631 430273 5.61 2.12

Example 31: 7-Day Toxicology Study in Mice Toxicology Summary

A toxicology study may be performed as described in this Example. Overtsigns of toxicity after 7 days of repeat dosing up to 100 mg/kg P.O. interms of clinical observations, body weight or food consumption will beexamined. White blood cells will be monitored and internal organsexamined after necroscopy.

TABLE 14 Toxicology Study Design Test system C57BL/6 Mouse, 5 weeks old,18-20 g, male, N = 12 Food status Free access to food and waterAdministration Group 1: 0 mg/kg/day (10 mL/kg/day) via oral gavage (N =3) Group 2: 10 mg/kg/day (10 mL/kg/day) via oral gavage (N = 3) Group 3:30 mg/kg/day (10 mL/kg/day) via oral gavage (N = 3) Group 4: 100mg/kg/day (10 mL/kg/day) via oral gavage (N = 3)

Toxicokinetics

Mean plasma, brain and CSF concentration-time profiles of test compoundswill be measured after a single PO administration at 30 mg/kg in maleC57BL/6 mice (5 weeks old) (N=3/time point). Mean plasma, brain and CSFconcentration-time profiles of compounds after repeat PO administrationsat 30 mg/kg in male C57BL/6 mice on day 7 (5 weeks old) (N=3/time point)will also be measured.

In-Life Summary:

The study design (36 animals, C57BL/6 mouse) consists of administratingthe drug [PO: 30 mg/kg/day (10 mL/kg/day) via oral gavage] andcollecting samples at terminal bleeding for plasma, brain and CSF at0.025, 0.5, 1, 4, 8 and 24 hr. The PO dosing solutions will be preparedin 50 mM citrate buffer (pH 4.0) at 3 mg/mL. The blood collection willbe performed as follows: the animal will be anesthetized underisoflurane. Approximately 500 μL blood/time point will be collected intoK₂EDTA tube via cardiac puncture for terminal bleeding. -200 μL bloodsamples will be put on ice and centrifuged to obtain a plasma sample(2000 g, 5 min under 4° C.) within 15 minutes of collection. -300 bloodsamples will be used for hematology assay. The brain collection will beperformed as follows: a mid-line incision will be made in the animal'sscalp and skin retracted. The skull overlying the brain will be removed.The whole brain will be collected, rinsed with cold saline, dried onfiltrate paper, weighted, and snap frozen by placing into dry ice. Thebrain sample will be homogenized for 2 min with 3 volumes of PBS (pH7.4) by Mini-bead-beater before sample extraction.

Plasma Samples Preparation:

An aliquot of 10 μL sample will be added to 200 μL MeCN containing 10ng/mL IS (Glipizide). The mixture will be vortexed for 10 min andcentrifuged at 6,000 rpm for 10 min. An aliquot of 1 μL constitutionwill be injected for LC-MS/MS analysis.

CSF Samples Preparation:

An aliquot of 3 μL sample will be added to 70 μL MeCN containing 10ng/mL IS (Glipizide). The mixture will be vortexed for 2 min andcentrifuged at 14,000 rpm for 5 min. An aliquot of 1 μL constitutionwill be injected for LC-MS/MS analysis.

Tissue Samples Preparation:

The sample will be homogenized with 3 volumes (v/w) of PBS. An aliquotof 10 μL sample will be added to 200 μL MeCN containing 10 ng/mL IS(Glipizide). The mixture will be vortexed for 10 min and centrifuged at6,000 rpm for 10 min. An aliquot of 1 μL constitution will be injectedfor LC-MS/MS analysis.

Analytical Method:

The sample analysis will be performed on LCMSMS-28 (Triple Quad 6500+)under the following conditions: positive ion, ESI, MRM detection usingglipizide as internal standard. HPLC conditions: mobile phase A:H₂O/0.025% FA with 1 mM NH₄₀Ac, mobile phase B: MeOH/0.025% FA with 1 mMNH₄₀Ac on Waters X-Bridge BEH C18 (2.1×50 mm, 2.5 μm) column at 60° C.

Example 32: Pharmacokinetics of Compounds After Intravenous or OralAdministration to Male Beagle Dogs

In-Life Summary:

The study design (9 animals, fasted overnight and fed at 4 h postdosing) consists of administrating the drug [IV: 1 mg/kg via cephalicvein injection], [PO: 3 mg/kg and 10 mg/kg via oral gavage] andcollecting samples at serial bleeding for plasma at 0.03, 0.08, 0.25,0.5, 1, 2, 4, 8, 24, 48 and 72 hr. The IV and PO dosing solutions wereprepared in 50 mM citrate buffer (pH 4.0) at 0.5 mg/mL, 1.5 mg/mL and 5mg/mL, respectively. The blood collection was performed as follows: theanimals were restrained manually, and approx. 0.5 mL blood/time pointwas collected from the cephalic vein into pre-cooled K₂EDTA tubes. Bloodsamples were put on wet ice and centrifuged at 4° C. to obtain plasmawithin 15 minutes of sample collection. All samples were stored atapproximately −70° C. until analysis.

Plasma Samples Preparation:

An aliquot of 30 μL sample was added to 100 MeCN containing 200 ng/mL IS(Dexamethasone). The mixture was vortexed for 10 min and centrifuged at5,800 rpm for 10 min. An aliquot of 30 μL supernatant was added to 60 μLH₂O and the mixture was vortexed for 5 min. An aliquot of 4 μLsupernatant was injected for LC-MS/MS analysis.

Analytical Method:

The sample analysis was performed using UPLC-MS/MS-02 (Triple Quad™4000) under the following conditions: positive ion, ESI, MRM detectionusing dexamethasone as internal standard. HPLC conditions: mobile phaseA: H₂O-0.1% FA, mobile phase B: ACN-0.1% FA on ACQUITY UPLC HSS T3(2.1×50 mm, 1.8 μm) column at 60° C.

TABLE 15 Pharmacokinetics of I-188 in Beagle Dogs Dose Cmax t_(1/2) ClVss AUC p Compound Species (mg/kg) (ng/mL) (hr) (L/hr/kg) (L/kg)(hr*ng/mL) % F I-188 dog IV: 1  — 27.3 1.19 39 722 — dog PO: 3  46.127.3 — — 1419 65 dog PO: 10 280 23.2 — — 7503 97.8

While we have described a number of embodiments of this invention, it isapparent that our basic examples may be altered to provide otherembodiments that utilize the compounds and methods of this invention.Therefore, it will be appreciated that the scope of this invention is tobe defined by the appended claims rather than by the specificembodiments that have been represented by way of example.

We claim:
 1. A method of treating a CXCR4-mediated disease, disorder, orcondition, comprising administering to a patient in need thereof acompound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein: Ring A is a 3-8membered saturated or partially unsaturated monocyclic carbocyclic ring,phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 4-8membered saturated or partially unsaturated monocyclic heterocyclic ringhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, orsulfur, a 5-6 membered monocyclic heteroaromatic ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur, oran 8-10 membered bicyclic heteroaromatic ring having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur; each R¹ isindependently —R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃, —SR, or -L¹-R⁶;each R is independently hydrogen or an optionally substituted groupselected from C₁₋₆ aliphatic, a 3-8 membered saturated or partiallyunsaturated monocyclic carbocyclic ring, phenyl, an 8-10 memberedbicyclic aromatic carbocyclic ring, a 4-8 membered saturated orpartially unsaturated monocyclic heterocyclic ring having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur, a5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or an 8-10membered bicyclic heteroaromatic ring having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur; each L¹ and L²is independently a covalent bond or a C₁₋₈ bivalent straight or branchedhydrocarbon chain wherein 1, 2, or 3 methylene units of the chain areindependently and optionally replaced with —O—, —C(O)—, —C(O)O—,—OC(O)—, —N(R)—, —C(O)N(R)—, —(R)NC(O)—, —OC(O)N(R)—, —(R)NC(O)O—,—N(R)C(O)N(R)—, —S—, —SO—, —SO₂—, —SO₂N(R)—, —(R)NSO₂—, —C(S)—, —C(S)O—,—OC(S)—, —C(S)N(R)—, —(R)NC(S)—, —(R)NC(S)N(R)—, or -Cy-; each -Cy- isindependently a bivalent optionally substituted 3-8 membered saturatedor partially unsaturated monocyclic carbocyclic ring, optionallysubstituted phenylene, an optionally substituted 4-8 membered saturatedor partially unsaturated monocyclic heterocyclic ring having 1-3heteroatoms independently selected from nitrogen, oxygen, or sulfur, anoptionally substituted 5-6 membered monocyclic heteroaromatic ringhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur, an optionally substituted 8-10 membered bicyclic or bridgedbicyclic saturated or partially unsaturated heterocyclic ring having 1-5heteroatoms independently selected from nitrogen, oxygen, or sulfur, oran optionally substituted 8-10 membered bicyclic or bridged bicyclicheteroaromatic ring having 1-5 heteroatoms independently selected fromnitrogen, oxygen, or sulfur; R² is hydrogen, halogen, —CN, —OR, —N(R)₂,—NO₂, —N₃, —SR, -L²-R⁶, or optionally substituted C₁₋₈ aliphatic; R³ ishydrogen, optionally substituted C₁₋₆ aliphatic, or -L³-R⁶; L³ is a C₁₋₆bivalent straight or branched hydrocarbon chain wherein 1, 2, or 3methylene units of the chain are independently and optionally replacedwith —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—, —C(O)N(R)—, —(R)NC(O)—, —S—,—SO—, —SO₂—, —C(S)—, or -Cy-; each R⁴ is independently hydrogen,deuterium, halogen, —CN, —OR⁶, or C₁₋₄ alkyl, or two R⁴ groups on thesame carbon are optionally taken together to form ═NR⁶, ═NOR⁶, ═O, or═S; each R⁵ is independently R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃,—SR, or -L¹-R⁶, or two R⁵ groups on the same saturated carbon atom areoptionally taken together to form ═NR, ═NOR, ═O, ═S, or a spirocyclic3-6 membered carbocyclic ring; each R⁶ is independently hydrogen or C₁₋₆alkyl optionally substituted with 1, 2, 3, 4, 5, or 6 deuterium orhalogen atoms; m is 0, 1, 2, 3, or 4; n is 0, 1, 2, 3, or 4; and p is 0,1, 2, 3, or
 4. 2. (canceled)
 3. The method of claim 1, wherein Ring A isselected from:


4. The method of claim 1, wherein Ring A is selected from


5. The method of claim 4, wherein R¹ is selected from hydrogen, halogen,C₁₋₆ alkyl (optionally substituted with 1, 2, or 3 halogens), —CN,—N(R)₂, —OR, —SR, —S(O)R⁶, —SO₂R⁶, —SO₂NHR⁶,

and each R is independently hydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl,cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, —CH₂F, —CHF₂, —CF₃,—CH₂CHF₂, or —CH₂CF₃.
 6. The method of claim 5, wherein L¹ is a C₁₋₆bivalent straight or branched hydrocarbon chain wherein 1, 2, or 3methylene units of the chain are independently and optionally replacedwith —O—, —C(O)—, —N(R)—, —S—, —SO—, —SO₂—, —SO₂N(R)—, —(R)NSO₂—,—C(S)—, or -Cy-, wherein each R is independently hydrogen, —CH₂-phenyl,phenyl, C₁₋₆ alkyl, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl,—CHF₂, —CF₃, —CH₂CHF₂, or —CH₂CF₃.
 7. The method of claim 6, wherein L²is a C₁₋₆ bivalent straight or branched hydrocarbon chain wherein 1, 2,or 3 methylene units of the chain are independently and optionallyreplaced with —O—, —C(O)—, —N(R)—, —S—, —SO—, —SO₂—, —SO₂N(R)—,—(R)NSO₂—, —C(S)—, or -Cy-, and each R is independently hydrogen,—CH₂-phenyl, phenyl, C₁₋₆ alkyl, cyclopropyl, cyclobutyl, cyclopentyl,cyclohexyl, —CF₃, —CH₂CHF₂, or —CH₂CF₃.
 8. (canceled)
 9. The method ofclaim 7, wherein R² is hydrogen, halogen, —CN, —OR, —N(R)₂, —SR,optionally substituted C₁₋₆ aliphatic, or -L²-R⁶, wherein L² is a C₁₋₆bivalent straight or branched hydrocarbon chain wherein 1, 2, or 3methylene units of the chain are independently and optionally replacedwith —O—, —C(O)—, —N(R)—, —S—, —SO—, —SO₂—, —C(S)—, or -Cy-; wherein theC₁₋₆ aliphatic group is optionally substituted with 1, 2, or 3 groupsindependently selected from halogen, —CN, —N(R)₂, —NO₂, —N₃, ═NR, ═NOR,═O, ═S, —OR, —SR, —SO₂R, —S(O)R, —R, -Cy-R, —C(O)R, —C(O)OR, —OC(O)R,—C(O)N(R)₂, —(R)NC(O)R, —OC(O)N(R)₂, —(R)NC(O)OR, —N(R)C(O)N(R)₂,—SO₂N(R)₂, —(R)NSO₂R, —C(S)R, or —C(S)OR; wherein each R isindependently hydrogen, —CH₂-phenyl, phenyl, C₁₋₆ alkyl, cyclopropyl,cyclobutyl, cyclopentyl, cyclohexyl, —CF₃, —CH₂CHF₂, or —CH₂CF₃. 10.(canceled)
 11. The method of claim 7, wherein R² is hydrogen, —NH₂, C₂₋₄alkynyl, F, Cl, Br, or I.
 12. (canceled)
 13. The method of claim 9,wherein R³ is C₁₋₄ alkyl optionally substituted with

pyridyl, —N(R)₂, —CN, or 1, 2, or 3 deuterium or halogen atoms, whereinR is hydrogen or C₁₋₃ alkyl.
 14. The method of claim 9, wherein R³ ismethyl.
 15. (canceled)
 16. The method of claim 14, wherein R⁵ ishydrogen, C₁₋₆ alkyl, halogen, —CN, —OCF₃, cyclopropyl, ethynyl, —OCH₃,—CF₃, —CD₃, or

17-21. (canceled)
 22. The method of claim 13, wherein the compound is ofFormula VII:

or a pharmaceutically acceptable salt thereof.
 23. The method of claim1, wherein the compound is of Formulae VIII-a or VIII-b:

or a pharmaceutically acceptable salt thereof. 24-28. (canceled)
 29. Themethod of claim 1, wherein the compound is of Formulae XIV-a, XIV-b, orXIV-c:

or a pharmaceutically acceptable salt thereof.
 30. The method of claim1, wherein the compound is selected from one of the following:

or a pharmaceutically acceptable salt thereof. 31-35. (canceled)
 36. Themethod of claim 1, wherein the CXCR4-mediated disease, disorder, orcondition is a cancer selected from a leukemia, polycythemia vera, alymphoma, Waldenstrom's macroglobulinemia, multiple myeloma, or heavychain disease; or a solid tumor selected from fibrosarcoma, myxosarcoma,liposarcoma, chondrosarcoma, osteogenic sarcoma, chordoma, angiosarcoma,endotheliosarcoma, lymphangiosarcoma, lymphangioendotheliosarcoma,synovioma, mesothelioma, Ewing's tumor, leiomyosarcoma,rhabdomyosarcoma, colon carcinoma, pancreatic cancer, breast cancer,ovarian cancer, prostate cancer, squamous cell carcinoma, basal cellcarcinoma, adenocarcinoma, sweat gland carcinoma, sebaceous glandcarcinoma, papillary carcinoma, papillary adenocarcinomas,cystadenocarcinoma, medullary carcinoma, bronchogenic carcinoma, renalcell carcinoma, hepatoma, bile duct carcinoma, choriocarcinoma,seminoma, embryonal carcinoma, Wilm's tumor, cervical cancer, uterinecancer, testicular cancer, lung carcinoma, small cell lung carcinoma,bladder carcinoma, or epithelial carcinoma.
 37. The method of claim 1,wherein the CXCR4-mediated disease, disorder, or condition is a leukemiaselected from acute leukemia, acute lymphocytic leukemia, acutemyelocytic leukemia, acute myeloblastic leukemia, acute promyelocyticleukemia, acute myelomonocytic leukemia, acute monocytic leukemia, acuteerythroleukemia, chronic leukemia, chronic myelocytic leukemia, orchronic lymphocytic leukemia; or a lymphoma selected from Hodgkin'sdisease or non-Hodgkin's disease.
 38. The method of claim 1, wherein theCXCR4-mediated disease, disorder, or condition is a cancer selected fromliver cancer, kidney cancer, ovarian cancer, ovarian epithelial cancer,fallopian tube cancer, papillary serous cystadenocarcinoma, uterinepapillary serous carcinoma (UPSC), colon cancer, prostate cancer,testicular cancer, rectal cancer, anal cancer, colon cancer, lungcancer, gallbladder cancer; hepatocholangiocarcinoma; soft tissue andbone synovial sarcoma; rhabdomyosarcoma; osteosarcoma; chondrosarcoma;Ewing sarcoma; anaplastic thyroid cancer; adrenocortical adenoma;pancreatic cancer; pancreatic ductal carcinoma or pancreaticadenocarcinoma; gastrointestinal/stomach (GIST) cancer; squamous cellcarcinoma of the head and neck (SCCHN); salivary gland cancer; orneurofibromatosis-1 associated malignant peripheral nerve sheath tumors(MPNST).
 39. The method of claim 1, wherein the CXCR4-mediated disease,disorder, or condition is a cancer selected from renal cell carcinoma,hepatocellular carcinoma (HCC), hepatoblastoma, melanoma, breast cancer,colorectal carcinoma, colorectal cancer, non-small cell lung cancer(NSCLC) or small cell lung cancer (SCLC), fallopian tube cancer,papillary serous cystadenocarcinoma, or uterine papillary serouscarcinoma (UPSC).
 40. The method of claim 1, wherein the CXCR4-mediateddisease, disorder, or condition is a primary immune deficiency selectedfrom: warts, hypogammaglobulinemia, infections, myelokathexis (WHIM)syndrome; severe congenital neutropenia (SCN); GATA2 deficiency (MonoMAC syndrome); idiopathic CD4+ T lymphocytopenia (ICL); orWiskott-Aldrich Syndrome.
 41. A method of inhibiting activity of CXCR4or a mutant thereof in a biological sample comprising the step ofcontacting the biological sample with a compound of Formula I:

or a pharmaceutically acceptable salt thereof, wherein: Ring A is a 3-8membered saturated or partially unsaturated monocyclic carbocyclic ring,phenyl, an 8-10 membered bicyclic aromatic carbocyclic ring, a 4-8membered saturated or partially unsaturated monocyclic heterocyclic ringhaving 1-2 heteroatoms independently selected from nitrogen, oxygen, orsulfur, a 5-6 membered monocyclic heteroaromatic ring having 1-4heteroatoms independently selected from nitrogen, oxygen, or sulfur, oran 8-10 membered bicyclic heteroaromatic ring having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur; each R¹ isindependently —R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃, —SR, or -L¹-R⁶;each R is independently hydrogen or an optionally substituted groupselected from C₁₋₆ aliphatic, a 3-8 membered saturated or partiallyunsaturated monocyclic carbocyclic ring, phenyl, an 8-10 memberedbicyclic aromatic carbocyclic ring, a 4-8 membered saturated orpartially unsaturated monocyclic heterocyclic ring having 1-2heteroatoms independently selected from nitrogen, oxygen, or sulfur, a5-6 membered monocyclic heteroaromatic ring having 1-4 heteroatomsindependently selected from nitrogen, oxygen, or sulfur, or an 8-10membered bicyclic heteroaromatic ring having 1-5 heteroatomsindependently selected from nitrogen, oxygen, or sulfur; each L¹ and L²is independently a covalent bond or a C₁₋₈ bivalent straight or branchedhydrocarbon chain wherein 1, 2, or 3 methylene units of the chain areindependently and optionally replaced with —O—, —C(O)—, —C(O)O—,—OC(O)—, —N(R)—, —C(O)N(R)—, —(R)NC(O)—, —OC(O)N(R)—, —(R)NC(O)O—,—N(R)C(O)N(R)—, —S—, —SO—, —SO₂—, —SO₂N(R)—, —(R)NSO₂—, —C(S)—, —C(S)O—,—OC(S)—, —C(S)N(R)—, —(R)NC(S)—, —(R)NC(S)N(R)—, or -Cy-; each -Cy- isindependently a bivalent optionally substituted 3-8 membered saturatedor partially unsaturated monocyclic carbocyclic ring, optionallysubstituted phenylene, an optionally substituted 4-8 membered saturatedor partially unsaturated monocyclic heterocyclic ring having 1-3heteroatoms independently selected from nitrogen, oxygen, or sulfur, anoptionally substituted 5-6 membered monocyclic heteroaromatic ringhaving 1-4 heteroatoms independently selected from nitrogen, oxygen, orsulfur, an optionally substituted 8-10 membered bicyclic or bridgedbicyclic saturated or partially unsaturated heterocyclic ring having 1-5heteroatoms independently selected from nitrogen, oxygen, or sulfur, oran optionally substituted 8-10 membered bicyclic or bridged bicyclicheteroaromatic ring having 1-5 heteroatoms independently selected fromnitrogen, oxygen, or sulfur; R² is hydrogen, halogen, —CN, —OR, —N(R)₂,—NO₂, —N₃, —SR, -L²-R⁶, or optionally substituted C₁₋₈ aliphatic; R³ ishydrogen, optionally substituted C₁₋₆ aliphatic, or -L³-R⁶; L³ is a C₁₋₆bivalent straight or branched hydrocarbon chain wherein 1, 2, or 3methylene units of the chain are independently and optionally replacedwith —O—, —C(O)—, —C(O)O—, —OC(O)—, —N(R)—, —C(O)N(R)—, —(R)NC(O)—, —S—,—SO—, —SO₂—, —C(S)—, or -Cy-; each R⁴ is independently hydrogen,deuterium, halogen, —CN, —OR⁶, or C₁₋₄ alkyl, or two R⁴ groups on thesame carbon are optionally taken together to form ═NR⁶, ═NOR⁶, ═O, or═S; each R⁵ is independently R, halogen, —CN, —OR, —N(R)₂, —NO₂, —N₃,—SR, or -L¹-R⁶, or two R⁵ groups on the same saturated carbon atom areoptionally taken together to form ═NR, ═NOR, ═O, ═S, or a spirocyclic3-6 membered carbocyclic ring; each R⁶ is independently hydrogen or C₁₋₆alkyl optionally substituted with 1, 2, 3, 4, 5, or 6 deuterium orhalogen atoms; m is 0, 1, 2, 3, or 4; n is 0, 1, 2, 3, or 4; and p is 0,1, 2, 3, or 4.